Abstract:
The present invention relates generally to the use of gene mutations, whose presence or absence are useful for predicting a patient&#39;s response to treatment with an anti-proliferative agent, in particular a WEE1 inhibitor. The presence or absence of a mutation to the TP53 gene, can be used to predict response to treatment with a WEE1 inhibitor in a patient presenting with a cancerous condition.

Description:
FIELD OF THE INVENTION 
       [0001]    The present invention relates generally to the use of gene mutations, whose presence or absence are useful for predicting a patient&#39;s response to treatment with an anti-proliferative agent, in particular a WEE1 inhibitor. The presence or absence of a mutation to the TP53 gene, can be used to predict response to treatment with a WEE1 inhibitor in a patient presenting with a cancerous condition. 
       BACKGROUND OF THE INVENTION 
       [0002]    Many commonly used anti-cancer drugs indiscriminately target DNA in dividing cells and ultimately cause DNA damage. This, in turn, triggers activation of cell cycle checkpoints which arrest progression of the cell cycle (at the G1, S, or G2/M phases) with the purpose of allowing time for the DNA to be repaired before the cell undergoes DNA replication or division. From a therapeutic standpoint, inhibition of checkpoint kinases that mediate cell cycle arrest could force tumor cells to continue cell division before chemically-induced DNA damage is repaired, eventually causing apoptosis or mitotic catastrophe (Medema, R. H. and Macurek, L.,  Oncogene,  2012, 31(21):2601-2613). Cell line studies support this hypothesis and show chemosensitization and radiosensitization by pharmacologic or genetic disruption of checkpoint kinase activity including CHK1, WEE1, ATR, and ATM. Inhibitors against these kinases are at various stages of preclinical and clinical development for their ability to sensitize tumor cells to therapeutic DNA damage. 
         [0003]    The checkpoint kinase WEE1 catalyzes an inhibitory phosphorylation of both CDK1 (CDC2) and CDK2 on tyrosine 15 (Parker, L. L. and Piwnica-Worms, H.,  Science,  1992, 257(5078):1955-1957; Watanabe, N., et al.,  Embo J.,  1995, 14(9):1878-1891). WEE1-dependent inhibition of CDK1 and CDK2 arrests the cell cycle in response to extrinsically induced DNA damage (Hamer, P. C. D., et al.,  Clin. Cancer Res.,  2011, 17(13):4200-4207). WEE1 activity is also essential for the unperturbed cell cycle (Mcgowan, C. H. and Russell, P.,  Embo J.,  1993, 12(1):75-85; Tominaga, Y., et al.,  Intl. J. Biol. Sci.,  2006, 2(4):161-170). Cell synchronization studies in normal human fibroblasts revealed that similar amounts of WEE1 protein were detected in both S and G2/M phases, but that its greatest activity was in S phase of the cell cycle (Watanabe, N., 1995). Further, upon conditional WEE1 knockout in mouse embryonic fibroblasts (MEFs), cells show evidence of genomic instability, malfunctioning checkpoints, and premature mitosis (Tominaga, et al., 2006). This phenotype was explained in part by recent findings that demonstrate a critical role for WEE1 in DNA synthesis. Knockdown of WEE1, in the absence of DNA damaging agents, led to rapid and robust detection of DNA double strand breaks specifically in S-phase cells undergoing DNA replication (Beck, H., et al.,  J. Cell Biol.,  2010, 188(5):629-638; Dominguez-Kelly, R., et al.,  J. Cell Biol.,  2011, 194(4):567-579). Data support a model of WEE1-dependent genomic stability in which WEE1 knockdown or inhibition leads to aberrantly high activity of CDK 1 and 2, resulting in inappropriately timed firing of excessive DNA replication origins that quickly depletes nucleotide pools and leads to stalled replication forks which, in the absence of WEE1 activity, are substrates for DNA exonucleases and resolve into DNA doubles strand breaks (Beck, H., et al., 2012). 
         [0004]    Deregulated WEE1 expression or activity is believed to be a hallmark of pathology in several types of cancer. WEE1 is often overexpressed in glioblastomas and its activity protects this tumor type from mitotic catastrophe such that high WEE1 levels are associated with poor prognosis (Mir, S. E., et al.,  Cancer Cell,  2010, 18(3):244-257). High expression of WEE1 was found in malignant melanoma and correlated with poor disease-free survival in this population (Magnussen, G. I., et al.,  Plos One,  2012, 7(6)). Aberrant WEE1 expression has been implicated in additional tumor types such as hepatocellular carcinoma (Masaki, T., et al.,  Hepatology,  2003, 37(3):534-543), breast cancer (Iorns, E., et al.,  Plos One,  2009, 4(4)), colon carcinoma (Backert, S., et al.,  Intl., J. Cancer,  1999, 82(6):868-874)), lung carcinoma (Yoshida, T., et al.,  Annals of Oncology,  2004, 15(2):252-256) and head and neck squamous cell carcinoma (Wu, Z. X., et al.,  Mol . &amp;  Cell. Proteomics,  2011, 10(12)). Advanced tumors with an increased level of genomic instability may require functional checkpoints to allow for repair of such lethal DNA damage. As such, WEE1 represents an attractive target in advanced tumors where its inhibition is believed to result in irreparable DNA damage (Sorensen, C. S. and Syljuasen, R. G.,  Nuc. Acids Res.,  2012, 40(2):477-486). 
         [0005]    The TP53 gene, which encodes the p53 protein, is an important regulator of the cell-cycle as a key regulator of the G 1  checkpoints and is one of the most frequently mutated genes in cancer (Molinari, M.,  Cell. Prolif.,  2000, 33:261-174). Cells lacking the G 1  checkpoint are predicted to be more dependent on the WEE1-mediated S or G2 checkpoint. Thus, p53-deficient tumors treated with G 2  checkpoint abrogators may be particularly susceptible to DNA damage (Kawabe, T.,  Mol. Cancer Ther.,  2004, 3:513-519; Bucher, N., and Britten, C. D.,  Br. J. Cancer,  2008, 98:523-528). 
         [0006]    There is a need for biomarkers that can be used to predict which patients are amenable to treatment with specific therapies, particularly for patients who are non-responsive or who are likely to become refractive to first line therapies. It is, therefore, an object of this invention to provide biomarkers to select patients likely to respond to treatment with a WEE1 inhibitor. 
       SUMMARY OF THE INVENTION 
       [0007]    The instant invention relates generally to the identification of TP53 gene mutations whose presence or absence are useful for evaluating and classifying patients for treatment with a WEE1 inhibitor. In one embodiment of the invention, the TP53 gene mutations resulting in loss of function are used to identify patients likely to respond to treatment with a WEE1 inhibitor. In another embodiment, the invention is a method for treating a patient diagnosed with a WEE1 associated cancer with a WEE1 inhibitor, wherein the cancer cells of said patient are characterized by the presence of a mutation in TP53 which renders p53 non-functional. In still another embodiment, the invention is a method for treating a cancer patient who is responsive or predicted to be responsive to treatment with a WEE1 inhibitor, wherein the cancer cells of said patient are characterized by the presence of a mutation in TP53 which renders p53 non-functional. 
     
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         [0008]      FIG. 1 : Effect of TP53 mutation status on the degree of synergy observed between WEE1-1 and various DNA-damaging agents. 
       
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
       [0009]    Many anti-cancer treatments act by damaging DNA, which subsequently initiates the DNA damage response (DDR) and activates checkpoint kinases to arrest division while the DNA is repaired. WEE1, a tyrosine kinase, is activated by the DDR to phosphorylate and inhibit cyclin dependent kinases (CDKs) 1 and 2 and, as such, arrest cell division. Inhibiting WEE1 potentiates DNA damaging treatments by abrogating cell cycle arrest and proper DNA repair. 
         [0010]    WEE1-1, also known as 2-allyl-1-[6-(1-hydroxy-1-methylethyl)pyridin-2-yl]-6-{[4-(4-methylpiperazin-1-yl)phenyl]amino}-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one, is a potent (IC 50 =5.2 nM) and selective ATP-competitive small molecule inhibitor of WEE1 (Hirai, H., et al.,  Mol. Cancer Ther.,  2009, 8(11):2992-3000) that is currently under clinical development as an anti-tumor agent in combination with standard of care (SOC) chemotherapeutics (Stathis, A. and Oza A.,  Drug News  &amp;  Perspectives,  2010, 23(7):425-429; Schellens, J. H. M., et al.,  J. Clin. Oncol.,  2011, 29:2011 (suppl; abstr 3068); Mizuarai, S., et al.,  Mol. Cancer,  2009, 8:34). Previous studies on WEE1-1 have demonstrated its potential as an adjunct or sensitizer to currently used standard of care (SOC) chemotherapeutics by its ability to force unscheduled mitosis that ultimately results in apoptosis or mitotic catastrophe (Hirai, H., et al.,  Cancer Biol . &amp;  Ther.,  2010, 9(7):514-522; Aarts, M., et al.,  Cancer Discovery,  2012, 2(6):524-539; Indovina, P. and Giordano A.,  Cancer Biol . &amp;  Ther.,  2010, 9(7); 523-525; Wang, Y. L., et al.,  Cancer Biol . &amp;  Ther.,  2004, 3(3):305-313). However, the potential therapeutic effect of WEE1 inhibition in the absence of SOC chemotherapy is less defined. RNAi knockdown of WEE1 inhibited proliferation of cancer cell lines (Iorns, E., et al.,  Cancer Targets,  2009 , Plos One,  4(4); Murrow, L. M., et al.,  Breast Cancer Research and Treatment,  2010, 122(2):347-357) and recently it was demonstrated that WEE1-1 alone can induce apoptosis in sarcoma cell lines treated in vitro (Kreahling, J. M., et al.,  Mol. Cancer Ther.,  2012, 11(1):174-182). 
         [0011]    The p53 protein is encoded by the TP53 gene. Pre-clinical studies have suggested that WEE1-1 may be selectively effective in patients having p53-defective tumors, that is, in patients whose tumors harbor mutations in TP53 which render p53 non-functional. While over 25,000 mutations in the TP53 gene have been reported, not every mutation is expected to lead to a loss of function. Moreover, even as to mutations associated with loss of function, all are not equal in their ability to extinguish the functionality of p53. 
         [0012]    Applicants herein have developed a method to identify a subset of loss of function mutations, referred to as a “p53 filter”, by assigning to each type of mutation (see Table 1) a point value related to the likelihood that the mutation results in loss of function of p53, and in one embodiment, sensitivity to treatment with a WEE1 inhibitor, such as WEE1-1. As shown in Table 3, the evidence score of each gene mutation was calculated based on adding together the point values for the mutation: stop codon mutation, splice site mutation, dominant negative mutation, p53 signature p value &lt;0.05, mutation reported &gt;10× times in somatic tissue at amino acid level, mutation reported &gt;10× in somatic tissue at nucleotide level. The TP53 mutation score of a patient is calculated by identifying the various gene mutations in Table 3 in the cancer cell of the patient, and adding together the evidence score of each identified gene mutation. 
         [0013]    Thus, in one embodiment of the invention, patients whose TP53 mutation score is 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0 or greater according to Table 3 are identified as those patients most likely to respond to treatment with a WEE1 inhibitor and are selected for treatment with a WEE1 inhibitor. In another embodiment of the invention, patients treated with a WEE1 inhibitor whose TP53 mutation score is 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0 or greater according to Table 3 are identified as patients most likely to continue to respond to treatment and are selected to continue treatment with a WEE1 inhibitor. 
         [0014]    The GENBANK accession number of the p53 protein is NM000546. The GENBANK accession number of the p53 gene is X54156.1. The TP53 gene sequence is also available from the IARC database (http://p53.iarc.fr). While different databases may use different nucleotide or amino acid numbering systems, based on the nucleotide and amino acid information in Table 3, one skilled in the art can readily identify the gene mutation and its location in the p53 gene. 
         [0015]    The present invention provides a method of treating cancer or modulating WEE1 activity in a patient comprising the step of:
       1) selecting a patient diagnosed with cancer that has one or more TP53 gene mutations according to Table 3, at least one basepair insertion or deletion in the TP53 gene that causes a frameshift in encoding the p53 protein resulting in loss of function, or a combination thereof in the cancer cell;   2) administering a therapeutically effective amount of a WEE1 inhibitor and optionally one or more additional anti-cancer agents to the patient.       
 
         [0018]    The present invention also provides a method of treating cancer or modulating WEE1 activity in a patient, in which the patient is diagnosed with cancer and has one or more TP53 gene mutations according to Table 3, at least one basepair insertion or deletion in the TP53 gene that causes a frameshift in encoding the p53 protein resulting in loss of function, or a combination thereof in the cancer cell; comprising the step of administering a therapeutically effective amount of a WEE1 inhibitor and optionally one or more additional anti-cancer agents to the patient. 
         [0019]    The present invention further provides a method of treating cancer or modulating WEE1 activity in a patient, comprising the step of administering a therapeutically effective amount of a WEE1 inhibitor and optionally one or more additional anti-cancer agents to the patient, wherein the patient is diagnosed with cancer and has one or more TP53 gene mutations according to Table 3, at least one basepair insertion or deletion in the TP53 gene that causes a frameshift in encoding the p53 protein resulting in loss of function, or a combination thereof in the cancer cell. In one embodiment, the invention provides a WEE1 inhibitor for use in the treatment of cancer or modulating WEE1 activity in a patient, wherein the patient is diagnosed with cancer and has one or more TP53 gene mutations according to Table 3, or at least one basepair insertion or deletion in the TP53 gene that causes a frameshift in encoding the p53 protein resulting in loss of function, or a combination thereof in the cancer cell, and the treatment optionally comprises one or more additional anti-cancer agents. 
         [0020]    In one embodiment, the at least one basepair insertion or deletion in the TP53 gene that causes a frameshift in encoding the p53 protein completely eliminates the DNA binding domain of p53. 
         [0021]    In one embodiment, the TP53 gene mutation has an evidence score that is equal or greater than 2.5 according to Table 3. In another embodiment, the TP53 gene mutation has an evidence score that is equal or greater than 3 according to Table 3. In another embodiment, the TP53 gene mutation has an evidence score that is equal or greater than 3.5 according to Table 3. In a further embodiment, the TP53 gene mutation has an evidence score that is equal or greater than 4.0 according to Table 3. In yet a further embodiment, the TP53 gene mutation has an evidence score that is equal or greater than 4.5 according to Table 3. In yet another embodiment, the TP53 gene mutation has an evidence score that is equal to 5.0 according to Table 3. In yet another embodiment, the patient has at least one of the TP53 gene mutations resulting in the amino acid change selected from the group consisting of C238F, R248W and R273L according to Table 3, a stop codon at the codon encoding E298 in the p53 protein, or a deletion of a basepair in the codon encoding V157 in the p53 protein. 
         [0022]    In another embodiment of the above method, step 1) is selecting a patient diagnosed with cancer that has two or more TP53 gene mutations according to Table 3, or three or more TP53 gene mutations according to Table 3 in the cancer cell. 
         [0023]    In one embodiment of the invention, the AmpliChip p53 Assay (Roche Molecular Systems, Inc., Pleasanton, Calif.) is used to identify the TP53 mutation present in the samples from patients diagnosed with a TP53 associated cancer. The AmpliChip p53 test is a microarray-based resequencing test. The test is designed to detect single nucleotide substitutions and lbp deletions in the entire coding region and the flanking splice sites of exons 2-11 of the TP53 gene in either formalin-fixed paraffin-embedded tissue (FFPE) or freshly frozen tissue. The AmpliChip p53 test queries for the presence of sequence alterations through comparative analysis of the hybridization pattern of a series of probes to sample DNA and wild-type reference DNA. The highly redundant probe tiling approach is able to detect a significantly lower abundance of TP53 mutations in samples which contain mixtures of normal and tumor tissue without the need for microdissection. See Li et al.  Current Genomics,  2008, 9, 466-474. 
         [0024]    Those skilled in the art would recognize and appreciate that other methods could be employed to identify the gene mutation present, such as Sanger sequencing, massively parallel sequencing (Next-Generation Sequencing), mass spectrometry, or PCR techniques. The inventive method herein is not tied to the method used to identify the specific p53 loss-of-function mutation or the absence of any such mutation. 
       WEE1 Inhibitors 
       [0025]    In an embodiment of the invention, the WEE1 inhibitor for use in the methods of the instant invention is WEE1-1, the structure of which is as shown below. 
         [0000]    
       
                 
         
             
             
         
       
     
         [0026]    WEE1-1 is a WEE1 inhibitor which is useful for the treatment of cancer. WEE1-1 is also known as 2-allyl-1-[6-(1-hydroxy-1-methylethyl)pyridin-2-yl]-6-{[4-(4-methylpiperazin-1-yl)phenyl]amino}-1,2-dihydro-3H-pyrazolo[3,4-d]pyrimidin-3-one. WEE1-1 has been described in U.S. Pat. No. 7,834,019, and in PCT International Publication WO2007/126122, WO 2007/126128 and WO2008/153207, which are incorporated by reference herein in their entirety. Crystalline forms of WEE1-1 are described in US Publication US2010-0124544 and PCT International Publication WO2011/034743, which are incorporated by reference herein in their entirety. 
         [0027]    In an embodiment of the invention, the WEE1 inhibitor for use in the instant invention is WEE1-2, the structure of which is as shown below. 
         [0000]    
       
                 
         
             
             
         
       
     
         [0028]    WEE1-2 is a WEE1 inhibitor which is useful for the treatment of cancer. WEE1-2 is also known as 3-(2,6-dichlorophenyl)-4-imino-7-[(2′-methyl-2′,3′-dihydro-1′H-spiro[cyclopropane-1,4′-isoquinolin]-7′-yl)amino]-3,4-dihydropyrimido[4,5-d]pyrimidin-2(1H)-one. WEE1-2 has been described in PCT International Publication WO2008/153207 and US Publication US2011-0135601, which are incorporated by reference herein in their entirety. Crystalline forms of WEE1-2 are described in International Publication WO2009/151997 and US Publication US2011-0092520. 
         [0029]    In one embodiment, the WEE1 inhibitor is 
         [0000]    
       
                 
         
             
             
         
       
     
         [0000]    or a pharmaceutically acceptable salt thereof.
 
In another embodiment, the WEE1 inhibitor is
 
         [0000]    
       
                 
         
             
             
         
       
     
         [0000]    or a pharmaceutically acceptable salt thereof. 
         [0030]    The compounds used in the methods of the present invention may have asymmetric centers, chiral axes, and chiral planes (as described in: E. L. Eliel and S. H. Wilen, Stereochemistry of Carbon Compounds, John Wiley &amp; Sons, New York, 1994, pages 1119-1190), and occur as racemates, racemic mixtures, and as individual diastereomers, with all possible isomers and mixtures thereof, including optical isomers, all such stereoisomers being included in the present invention. In addition, the compounds disclosed herein may exist as tautomers and both tautomeric forms are intended to be encompassed by the scope of the invention, even though only one tautomeric structure is depicted. 
         [0031]    In the compounds used in the methods of the present invention, the atoms may exhibit their natural isotopic abundances, or one or more of the atoms may be artificially enriched in a particular isotope having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number predominantly found in nature. The present invention is meant to include all suitable isotopic variations of the compounds disclosed herein. For example, different isotopic forms of hydrogen (H) include protium (1H) and deuterium (2H). Protium is the predominant hydrogen isotope found in nature. Enriching for deuterium may afford certain therapeutic advantages, such as increasing in vivo half-life or reducing dosage requirements, or may provide a compound useful as a standard for characterization of biological samples. Isotopically-enriched compounds disclosed herein can be prepared without undue experimentation by conventional techniques well known to those skilled in the art. 
         [0032]    The WEE1 inhibitors used in the methods of the instant invention may also exist as various crystals, amorphous substances, pharmaceutically acceptable salts, hydrates and solvates. Further, the WEE1 inhibitors of the instant invention may be provided as prodrugs. In general, such prodrugs are functional derivatives of the WEE1 inhibitors of the instant invention that can be readily converted into compounds that are needed by living bodies. Accordingly, in the method of treatment of various cancers in the invention, the term “administration” includes not only the administration of a specific compound but also the administration of a compound which, after administered to patients, can be converted into the specific compound in the living bodies. Conventional methods for selection and production of suitable prodrug derivatives are described, for example, in “Design of Prodrugs”, ed. H. Bundgaard, Elsevier, 1985, which is referred to herein and is entirely incorporated herein as a part of the present description. Metabolites of the compound may include active compounds that are produced by putting the compound in a biological environment, and are within the scope of the compound in the invention. 
         [0033]    In one embodiment of the invention, the WEE1 inhibitor is administered in a dose between 100 mg per day and 250 mg per day. In another embodiment of the invention, the WEE1 inhibitors may be dosed twice a day (BID) over the course of two and a half days (for a total of 5 doses) or once a day (QD) over the course of two days (for a total of 2 doses). 
         [0034]    In another embodiment of the invention, the WEE1 inhibitor is administered in a dose between 200 mg per day and 400 mg per day, and preferably 250-350 mg per day. In an embodiment of the invention, the WEE1 inhibitors may be dosed once a day (QD) over the course of five days. 
       Method of Treating Cancer 
       [0035]    Cancers that may be treated by the WEE1 inhibitors include, but are not limited to: Cardiac: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma and teratoma; Lung: bronchogenic carcinoma (squamous cell, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma; Gastrointestinal: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumors, vipoma), small bowel (adenocarcinoma, lymphoma, carcinoid tumors, Karposi&#39;s sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large bowel (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma) colorectal; Genitourinary tract: kidney (adenocarcinoma, Wilm&#39;s tumor [nephroblastoma], lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma); Liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; Bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing&#39;s sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, osteochronfroma (osteocartilaginous exostoses), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumors; Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma, osteitis deformans), meninges (meningioma, meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germinoma [pinealoma], glioblastoma multiform, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal cord neurofibroma, meningioma, glioma, sarcoma); Gynecological: uterus (endometrial carcinoma), cervix (cervical carcinoma, pre tumor cervical dysplasia), ovaries (ovarian carcinoma [serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma], granulosa thecal cell tumors, Sertoli-Leydig cell tumors, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal rhabdomyosarcoma), fallopian tubes (carcinoma), breast; Hematologic: blood (myeloid leukemia [acute and chronic], acute lymphoblastic leukemia, chronic lymphocytic leukemia, myeloproliferative diseases, multiple myeloma, myelodysplastic syndrome), Hodgkin&#39;s disease, non-Hodgkin&#39;s lymphoma [malignant lymphoma]; Skin: malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Karposi&#39;s sarcoma, moles dysplastic nevi, lipoma, angioma, dermatofibroma, keloids, psoriasis; and Adrenal glands: neuroblastoma. 
         [0036]    The term “WEE1 kinase associated cancer” as referred to in this description means a cancer associated with the activity or inhibition of WEE1 kinases including, but not limited to, brain cancer, cervicocerebral cancer, esophageal cancer, thyroid cancer, small cell cancer, non-small cell cancer, breast cancer, lung cancer, stomach cancer, gallbladder/bile duct cancer, liver cancer, pancreatic cancer, colon cancer, rectal cancer, ovarian cancer, choriocarcinoma, uterus body cancer, uterocervical cancer, renal pelvis/ureter cancer, bladder cancer, prostate cancer, penis cancer, testicles cancer, fetal cancer, Wilms&#39; cancer, skin cancer, malignant melanoma, neuroblastoma, osteosarcoma, Ewing&#39;s tumor, soft part sarcoma, acute leukemia, chronic lymphatic leukemia, chronic myelocytic leukemia, Hodgkin&#39;s lymphoma, or as sensitizers for chemo therapy or radiation therapy of those cancers. In particular, the WEE1 inhibitor of the invention are useful as remedies, for example, for breast cancer, lung cancer, pancreatic cancer, colon cancer, ovarian cancer, acute leukemia, chronic lymphatic leukemia, chronic myelocytic leukemia, Hodgkin&#39;s lymphoma, or as sensitizers for chemotherapy or radiation therapy of those cancers. 
         [0037]    In another embodiment, the cancer is selected from the group consisting of ovarian cancer, melanoma, lung cancer, colorectal cancer, colon cancer, rectum cancer, prostate cancer, and breast cancer. 
         [0038]    In a further embodiment, the cancer is selected from the group consisting of ovarian carcinoma, ovary clear cell carcinoma, ovary adenocarcinoma, ovary teratocarcinoma, skin malignant melanoma, malignant melanoma, lung carcinoma, large cell lung cancer, lung adenocarcinoma, non-small cell lung cancer, lung squamous cell carcinoma, colorectal carcinoma, colorectal adenocarcinoma, colon carcinoma, rectum adenocarcinoma, prostate carcinoma, breast ductal carcinoma and breast cancer. 
         [0039]    In yet another embodiment, the cancer is ovarian cancer. In a further embodiment, the cancer is lung cancer. 
         [0040]    In another embodiment of the invention, a method of inhibiting or modulating WEE1 activity in a patient is provided. 
         [0041]    The term “treatment of cancer” as referred to in this description means that an anti-cancer agent is administered to a cancer patient so as to inhibit the growth of the cancer cells in the patient. 
         [0042]    The term “patient” or “subject” as referred to in this description means the recipient in need of medical intervention or treatment. Mammalian and non-mammalian patients or subjects are included. 
       Combination Therapy 
       [0043]    Combinations of the WEE1 inhibitors with therapeutic, chemotherapeutic and anti-cancer agents in the methods of the invention are within the scope of the invention. The WEE1 inhibitors may also be administered in combination with one or more additional anti-cancer agents, wherein the amounts of the WEE1 inhibitor and the anti-cancer agent result in a therapeutic effect. Examples of such agents can be found in  Cancer Principles and Practice of Oncology  by V. T. Devita and S. Hellman (editors), 6 th  edition (Feb. 15, 2001), Lippincott Williams &amp; Wilkins Publishers. A person of ordinary skill in the art would be able to discern which combinations of agents would be useful based on the particular characteristics of the drugs and the cancer involved. Such agents include the following: estrogen receptor modulators, androgen receptor modulators, retinoid receptor modulators, cytotoxic/cytostatic agents, antiproliferative agents, prenyl-protein transferase inhibitors, HMG-CoA reductase inhibitors and other angiogenesis inhibitors, HIV protease inhibitors, reverse transcriptase inhibitors, inhibitors of cell proliferation and survival signaling, bisphosphonates, aromatase inhibitors, siRNA therapeutics, γ-secretase inhibitors, agents that interfere with receptor tyrosine kinases (RTKs) and agents that interfere with cell cycle checkpoints. The WEE1 inhibitors may also be useful when co-administered with radiation therapy. The WEE1 inhibitors can be present in the same dosage unit as the anticancer agent or in separate dosage units. 
         [0044]    Non-limiting examples of suitable anti-cancer agents include cytostatic agents, cytotoxic agents, targeted therapeutic agents (small molecules, biologics, siRNA and microRNA) against cancer and neoplastic diseases as follows:
       1) anti-metabolites (such as methoxtrexate, 5-fluorouracil, gemcitabine, fludarabine, capecitabine);   2) alkylating agents, such as temozolomide and cyclophosphamide,   3) DNA interactive and DNA damaging agents, such as cisplatin, oxaliplatin and doxorubicin,   4) Ionizing irradiation, such as radiation therapy,   5) topoisomerase II inhibitors, such as etoposide and doxorubicin,   6) topoisomerase I inhibitors, such as irinotecan and topotecan,   7) tubulin interacting agents, such as paclitaxel, docetaxel, abraxane and epothilones,   8) kinesin spindle protein inhibitors,   9) spindle checkpoint inhibitors,   10) poly(ADP-ribose) polymerase (PARP) inhibitors, such as olaparib, MK-4827 and veliparib,   11) matrix metalloprotease (MMP) inhibitors,   12) protease inhibitors, such as cathepsin D and cathepsin K inhibitors,   13) proteosome or ubiquitination inhibitors, such as bortezomib,   14) activators of mutant p53 to restore its wild-type p53 activity,   15) adenoviral-p53,   16) Bcl-2 inhibitors, such as ABT-263,   17) heat shock protein (HSP) modulators, such as geldanamycin and 17-AAG,   18) histone deacetylase (HDAC) inhibitors, such as vorinostat (SAHA),   19) sex hormone modulating agents,
           a. anti-estrogens, such as tamoxifen and fulvestrant,   b. selective estrogen receptor modulators (SERM), such as raloxifene,   c. anti-androgens, such as bicalutamide and flutamide,   d. LHRH agonists, such as leuprolide,   e. 5α-reductase inhibitors, such as finasteride,   f. cytochrome P450 C17 lyase (CYP450c17, also called 17α-hydroxylase/17,20 lysase) inhibitors, such as abiraterone acetate, VN/124-1 and TAK-700,   g. aromatase inhibitors, such as letrozole, anastrozole and exemestane,   
           20) EGFR kinase inhibitors, such as geftinib, erlotinib and laptinib,   21) dual erbB1 and erbB2 inhibitors, such as lapatinib,   22) multi-targeted kinases (serine/threonine and/or tyrosine kinase) inhibitors,   a. ABL kinase inhibitors, imatinib, nilotinib and dasatinib,
           b. VEGFR-1, VEGFR-2, PDGFR, KDR, FLT, c-Kit, Tie2, Raf, MEK and ERK inhibitors, such as sunitinib, sorafenib, vandetanib, pazopanib, PLX-4032, axitinib, PTK787 and GSK-1120212,   c. polo-like kinase inhibitors,   d. aurora kinase inhibitors,   e. JAK inhibitors,   f. c-MET kinase inhibitors,   g. cyclin-dependent kinase inhibitors, such as CDK1 and CDK2 inhibitor SCH 727965,   h. PI3K and mTOR inhibitors, such as GDC-0941, BEZ-235, BKM-120 and AZD-8055,   i. rapamycin and its analogs, such as temsirolimus, everolimus, and deforolimus   
           23) and other anti-cancer (also know as anti-neoplastic) agents include but are not limited to ara-C, adriamycin, cytoxan, carboplatin, uracil mustard, clormethine, ifosfsmide, melphalan, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, pentostatine, vinblastine, vincristine, vindesine, vinorelbine, navelbine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, teniposide, cytarabine, pemetrexed, idarubicin, mithramycin, deoxycoformycin, mitomycin-C, 1-asparaginase, teniposide, ethinylestradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate, testolactone, megestrolacetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone, aminoglutethimide, estramustine, flutamide medroxyprogesteroneacetate, toremifene, goserelin, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxantrone, levamisole, drolloxafine, hexamethylmelamine, bexxar, zevalin, trisenox, profimer, thiotepa, altretamine, doxil, ontak, depocyt, aranesp, neupogen, neulasta and kepivance,   24) farnesyl protein transferase inhibitors, such as, SARASAR™ (4-[2-[4-[(11R)-3,10-dibromo-8-chloro-6, 11-dihydro-5H-benzo[5,6]cyclohepta[1,2-b]pyridin-11-yl-]-1-piperidinyl]-2-oxoethyl]-piperidinecarboxamide and tipifarnib,   25) interferons, such as Intron A and Peg-Intron,   26) anti-erbB1 antibodies, such as cetuximab and panitumumab,   27) anti-erbB2 antibodies, such as trastuzumab,   28) anti-CD52 antibodies, such as alemtuzumab,   29) anti-CD20 antibodies, such as rituximab,   30) anti-CD33 antibodies, such as gemtuzumab ozogamicin,   31) anti-VEGF antibodies, such as avastin,   32) TRIAL ligands, such as lexatumumab, mapatumumab, and AMG-655,   33) anti-CTLA-4 antibodies, such as ipilimumab,   34) antibodies against CTA1, CEA, CD5, CD19, CD22, CD30, CD44, CD44V6, CD55, CD56, EpCAM, FAP, MHCII, HGF, IL-6, MUC1, PSMA, TALE, TAG-72, TRAILR, VEGFR, IGF-2 and FGF, and   35) anti-IGF-1R antibodies, such as dalotuzumab (MK-0646) and robatumumab (SCH 717454).       
 
         [0096]    If formulated as a fixed dose such combination products employ the WEE1 inhibitor administered in the invention within the dosage range described herein and the other pharmaceutically active agent or treatment within its dosage range. The WEE1 inhibitor may also be administered sequentially with known anticancer or cytotoxic agents when a combination formulation is inappropriate. The invention is not limited in the sequence of administration; the WEE1 inhibitor may be administered either concurrent with, prior to or after administration of the known anticancer or cytotoxic agent. Such techniques are within the skills of the persons skilled in the art as well as attending physicians. 
         [0097]    Accordingly, in an aspect, this invention includes combinations for use in the invention comprising an amount of a WEE1 inhibitor, or a pharmaceutically acceptable salt thereof, and an amount of one or more anti-cancer treatments and anti-cancer agents listed above or below wherein the amounts of the compounds/treatments result in potential therapeutic effect. In one embodiment, the anti-cancer agent is selected from the group consisting of: 5-FU, carboplatin, doxorubicin, etoposide, gemcitabine, irinotecan, mitomycin, temozolmide and topotecan. In another embodiment, the anti-cancer agent is carboplatin. In another embodiment, the anti-cancer agents are carboplatin and paclitaxel. 
         [0098]    “Estrogen receptor modulators” refers to compounds that interfere with or inhibit the binding of estrogen to the receptor, regardless of mechanism. Examples of estrogen receptor modulators include, but are not limited to, tamoxifen, raloxifene, idoxifene, LY353381, LY117081, toremifene, fulvestrant, 4-[7-(2,2-dimethyl-1-oxopropoxy-4-methyl-2-[4-[2-(1-piperidinyl)ethoxy]phenyl]-2H-1-benzopyran-3-yl]-phenyl-2,2-dimethylpropanoate, 4,4′-dihydroxybenzophenone-2,4-dinitrophenyl-hydrazone, and SH646. 
         [0099]    “Androgen receptor modulators” refers to compounds which interfere or inhibit the binding of androgens to the receptor, regardless of mechanism. Examples of androgen receptor modulators include finasteride and other 5α-reductase inhibitors, nilutamide, flutamide, bicalutamide, liarozole, and abiraterone acetate. 
         [0100]    “Retinoid receptor modulators” refers to compounds which interfere or inhibit the binding of retinoids to the receptor, regardless of mechanism. Examples of such retinoid receptor modulators include bexarotene, tretinoin, 13-cis-retinoic acid, 9-cis-retinoic acid, α-difluoromethylornithine, ILX23-7553, trans-N-(4′-hydroxyphenyl) retinamide, and N-4-carboxyphenyl retinamide. 
         [0101]    “Cytotoxic/cytostatic agents” refer to compounds which cause cell death or inhibit cell proliferation primarily by interfering directly with the cell&#39;s functioning or inhibit or interfere with cell mitosis, including alkylating agents, tumor necrosis factors, intercalators, hypoxia activatable compounds, microtubule inhibitors/microtubule-stabilizing agents, inhibitors of mitotic kinesins, histone deacetylase inhibitors, inhibitors of kinases involved in mitotic progression, inhibitors of kinases involved in growth factor and cytokine signal transduction pathways, antimetabolites, biological response modifiers, hormonal/anti-hormonal therapeutic agents, haematopoietic growth factors, monoclonal antibody targeted therapeutic agents, topoisomerase inhibitors, proteosome inhibitors, ubiquitin ligase inhibitors, and aurora kinase inhibitors. 
         [0102]    Examples of cytotoxic/cytostatic agents include, but are not limited to, platinum coordinator compounds, sertenef, cachectin, ifosfamide, tasonermin, lonidamine, carboplatin, altretamine, prednimustine, dibromodulcitol, ranimustine, fotemustine, nedaplatin, oxaliplatin, temozolomide, heptaplatin, estramustine, improsulfan tosilate, trofosfamide, nimustine, dibrospidium chloride, pumitepa, lobaplatin, satraplatin, profiromycin, cisplatin, irofulven, dexifosfamide, cis-aminedichloro(2-methyl-pyridine)platinum, benzylguanine, glufosfamide, GPX100, (trans, trans, trans)-bis-mu-(hexane-1,6-diamine)-mu-[diamine-platinum(II)]bis[diamine(chloro)platinum (II)]-tetrachloride, diarizidinylspermine, arsenic trioxide, 1-(11-dodecylamino-10-hydroxyundecyl)-3,7-dimethylxanthine, zorubicin, idarubicin, daunorubicin, bisantrene, mitoxantrone, pirarubicin, pinafide, valrubicin, amrubicin, antineoplaston, 3′-deamino-3′-morpholino-13-deoxo-10-hydroxycarminomycin, annamycin, galarubicin, elinafide, MEN10755, 4-demethoxy-3-deamino-3-aziridinyl-4-methylsulphonyl-daunorubicin (see WO 00/50032). 
         [0103]    Examples of proteosome inhibitors include but are not limited to lactacystin and MLN-341 (Velcade). 
         [0104]    Examples of microtubule inhibitors/microtubule-stabilising agents include taxanes in general. Specific compounds include paclitaxel (Taxol®), vindesine sulfate, 3′,4′-didehydro-4′-deoxy-8′-norvincaleukoblastine, docetaxol (Taxotere®), rhizoxin, dolastatin, mivobulin isethionate, auristatin, cemadotin, RPR109881, BMS184476, vinflunine, cryptophycin, 2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl) benzene sulfonamide, anhydrovinblastine, N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-prolyl-L-proline-t-butylamide, TDX258, the epothilones (see for example U.S. Pat. Nos. 6,284,781 and 6,288,237) and BMS188797. 
         [0105]    Some examples of topoisomerase inhibitors are topotecan, hycaptamine, irinotecan, rubitecan, 6-ethoxypropionyl-3′,4′-O-exo-benzylidene-chartreusin, 9-methoxy-N,N-dimethyl-5-nitropyrazolo[3,4,5-kl]acridine-2-(6H) propanamine, 1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H,12H-benzo[de]pyrano[3′,4′:b,7]-indolizino[1,2b]quinoline-10,13(9H,15H)dione, lurtotecan, 7-[2-(N-isopropylamino)ethyl]-(20S)camptothecin, BNP1350, BNPI1100, BN80915, BN80942, etoposide phosphate, teniposide, sobuzoxane, 2′-dimethylamino-2′-deoxy-etoposide, GL331, N-[2-(dimethylamino)ethyl]-9-hydroxy-5,6-dimethyl-6H-pyrido[4,3-b]carbazole-1-carboxamide, asulacrine, (5a, 5aB, 8aa,9b)-9-[2-[N-[2-(dimethylamino)ethyl]-N-methylamino]ethyl]-5-[4-hydro0xy-3,5-dimethoxyphenyl]-5,5a,6,8,8a,9-hexohydrofuro(3′,4′:6,7)naphtho(2,3-d)-1,3-dioxol-6-one, 2,3-(methylenedioxy)-5-methyl-7-hydroxy-8-methoxybenzo[c]-phenanthridinium, 6,9-bis[(2-aminoethyl)amino]benzo[g]isoguinoline-5,10-dione, 5-(3-aminopropylamino)-7,10-dihydroxy-2-(2-hydroxyethylaminomethyl)-6H-pyrazolo[4,5,1-de]acridin-6-one, N-[1-[2(diethylamino)ethylamino]-7-methoxy-9-oxo-9H-thioxanthen-4-ylmethyl]formamide, N-(2-(dimethylamino)ethyl)acridine-4-carboxamide, 6-[[2-(dimethylamino)ethyl]amino]-3-hydroxy-7H-indeno[2,1-c] quinolin-7-one, and dimesna. 
         [0106]    Examples of inhibitors of mitotic kinesins, and in particular the human mitotic kinesin KSP, are described in Publications WO03/039460, WO03/050064, WO03/050122, WO03/049527, WO03/049679, WO03/049678, WO04/039774, WO03/079973, WO03/099211, WO03/105855, WO03/106417, WO04/037171, WO04/058148, WO04/058700, WO04/126699, WO05/018638, WO05/019206, WO05/019205, WO05/018547, WO05/017190, US2005/0176776. In an embodiment inhibitors of mitotic kinesins include, but are not limited to inhibitors of KSP, inhibitors of MKLP1, inhibitors of CENP-E, inhibitors of MCAK and inhibitors of Rab6-KIFL. 
         [0107]    Examples of “histone deacetylase inhibitors” include, but are not limited to, SAHA, TSA, oxamflatin, PXD101, MG98 and scriptaid. Further reference to other histone deacetylase inhibitors may be found in the following manuscript; Miller, T. A. et al.  J. Med. Chem.  46(24):5097-5116 (2003). 
         [0108]    “Inhibitors of kinases involved in mitotic progression” include, but are not limited to, inhibitors of aurora kinase, inhibitors of Polo-like kinases (PLK; in particular inhibitors of PLK-1), inhibitors of bub-1 and inhibitors of bub-R1. An example of an “aurora kinase inhibitor” is VX-680. 
         [0109]    “Antiproliferative agents” includes antisense RNA and DNA oligonucleotides such as G3139, ODN698, RVASKRAS, GEM231, and INX3001, and antimetabolites such as enocitabine, carmofur, tegafur, pentostatin, doxifluridine, trimetrexate, fludarabine, capecitabine, galocitabine, cytarabine ocfosfate, fosteabine sodium hydrate, raltitrexed, paltitrexid, emitefur, tiazofurin, decitabine, nolatrexed, pemetrexed, nelzarabine, 2′-deoxy-2′-methylidenecytidine, 2′-fluoromethylene-2′-deoxycytidine, N-[5-(2,3-dihydro-benzofuryl)sulfonyl]-N′-(3,4-dichlorophenyl)urea, N6-[4-deoxy-4-[N2-[2(E),4(E)-tetradecadienoyl]glycylamino]-L-glycero-B-L-manno-heptopyranosyl]adenine, aplidine, ecteinascidin, troxacitabine, 4-[2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimidino[5,4-b][1,4]thiazin-6-yl-(S)-ethyl]-2,5-thienoyl-L-glutamic acid, aminopterin, 5-flurouracil, alanosine, 11-acetyl-8-(carbamoyloxymethyl)-4-formyl-6-methoxy-14-oxa-1,11-diazatetracyclo(7.4.1.0.0)-tetradeca-2,4,6-trien-9-yl acetic acid ester, swainsonine, lometrexol, dexrazoxane, methioninase, 2′-cyano-2′-deoxy-N4-palmitoyl-1-B-D-arabin furanosyl cytosine, 3-aminopyridine-2-carboxaldehyde thiosemicarbazone and trastuzumab. 
         [0110]    Examples of monoclonal antibody targeted therapeutic agents include those therapeutic agents which have cytotoxic agents or radioisotopes attached to a cancer cell specific or target cell specific monoclonal antibody. Examples include Bexxar. 
         [0111]    “HMG-CoA reductase inhibitors” refers to inhibitors of 3-hydroxy-3-methylglutaryl-CoA reductase. Examples of HMG-CoA reductase inhibitors that may be used include but are not limited to lovastatin (MEVACOR®; see U.S. Pat. Nos. 4,231,938, 4,294,926 and 4,319,039), simvastatin (ZOCOR®; see U.S. Pat. Nos. 4,444,784, 4,820,850 and 4,916,239), pravastatin (PRAVACHOL®; see U.S. Pat. Nos. 4,346,227, 4,537,859, 4,410,629, 5,030,447 and 5,180,589), fluvastatin (LESCOL®; see U.S. Pat. Nos. 5,354,772, 4,911,165, 4,929,437, 5,189,164, 5,118,853, 5,290,946 and 5,356,896), atorvastatin (LIPITOR®; see U.S. Pat. Nos. 5,273,995, 4,681,893, 5,489,691 and 5,342,952) and cerivastatin (also known as rivastatin and BAYCHOL®; see U.S. Pat. No. 5,177,080). The structural formulas of these and additional HMG-CoA reductase inhibitors that may be used in the instant methods are described at page 87 of M. Yalpani, “Cholesterol Lowering Drugs”, Chemistry &amp; Industry, pp. 85-89 (5 Feb. 1996) and U.S. Pat. Nos. 4,782,084 and 4,885,314. The term HMG-CoA reductase inhibitor as used herein includes all pharmaceutically acceptable lactone and open-acid forms (i.e., where the lactone ring is opened to form the free acid) as well as salt and ester forms of compounds which have HMG-CoA reductase inhibitory activity, and therefor the use of such salts, esters, open-acid and lactone forms is included within the scope of this invention. 
         [0112]    “Prenyl-protein transferase inhibitor” refers to a compound which inhibits any one or any combination of the prenyl-protein transferase enzymes, including farnesyl-protein transferase (FPTase), geranylgeranyl-protein transferase type I (GGPTase-I), and geranylgeranyl-protein transferase type-II (GGPTase-II, also called Rab GGPTase). 
         [0113]    Examples of prenyl-protein transferase inhibitors can be found in the following publications and patents: WO 96/30343, WO 97/18813, WO 97/21701, WO 97/23478, WO 97/38665, WO 98/28980, WO 98/29119, WO 95/32987, U.S. Pat. No. 5,420,245, U.S. Pat. No. 5,523,430, U.S. Pat. No. 5,532,359, U.S. Pat. No. 5,510,510, U.S. Pat. No. 5,589,485, U.S. Pat. No. 5,602,098, European Patent Publ. 0 618 221, European Patent Publ. 0 675 112, European Patent Publ. 0 604 181, European Patent Publ. 0 696 593, WO 94/19357, WO 95/08542, WO 95/11917, WO 95/12612, WO 95/12572, WO 95/10514, U.S. Pat. No. 5,661,152, WO 95/10515, WO 95/10516, WO 95/24612, WO 95/34535, WO 95/25086, WO 96/05529, WO 96/06138, WO 96/06193, WO 96/16443, WO 96/21701, WO 96/21456, WO 96/22278, WO 96/24611, WO 96/24612, WO 96/05168, WO 96/05169, WO 96/00736, U.S. Pat. No. 5,571,792, WO 96/17861, WO 96/33159, WO 96/34850, WO 96/34851, WO 96/30017, WO 96/30018, WO 96/30362, WO 96/30363, WO 96/31111, WO 96/31477, WO 96/31478, WO 96/31501, WO 97/00252, WO 97/03047, WO 97/03050, WO 97/04785, WO 97/02920, WO 97/17070, WO 97/23478, WO 97/26246, WO 97/30053, WO 97/44350, WO 98/02436, and U.S. Pat. No. 5,532,359. For an example of the role of a prenyl-protein transferase inhibitor on angiogenesis see  European J. of Cancer , Vol. 35, No. 9, pp. 1394-1401 (1999). 
         [0114]    “Angiogenesis inhibitors” refers to compounds that inhibit the formation of new blood vessels, regardless of mechanism. Examples of angiogenesis inhibitors include, but are not limited to, tyrosine kinase inhibitors, such as inhibitors of the tyrosine kinase receptors Flt-1 (VEGFR1) and Flk-1/KDR (VEGFR2), inhibitors of epidermal-derived, fibroblast-derived, or platelet derived growth factors, MMP (matrix metalloprotease) inhibitors, integrin blockers, interferon-α, interleukin-12, pentosan polysulfate, cyclooxygenase inhibitors, including nonsteroidal anti-inflammatories (NSAIDs) like aspirin and ibuprofen as well as selective cyclooxy-genase-2 inhibitors like celecoxib and rofecoxib ( PNAS , Vol. 89, p. 7384 (1992);  JNCI, Vol.  69 , p.  475 (1982);  Arch. Opthalmol ., Vol. 108, p. 573 (1990);  Anat. Rec ., Vol. 238, p. 68 (1994);  FEBS Letters , Vol. 372, p. 83 (1995);  Clin, Orthop . Vol. 313, p. 76 (1995);  J. Mol. Endocrinol ., Vol. 16, p. 107 (1996);  Jpn. J. Pharmacol ., Vol. 75, p. 105 (1997);  Cancer Res ., Vol. 57, p. 1625 (1997);  Cell, Vol.  93 , p.  705 (1998);  Intl. J. Mol. Med ., Vol. 2, p. 715 (1998);  J. Biol. Chem ., Vol. 274, p. 9116 (1999)), steroidal anti-inflammatories (such as corticosteroids, mineralocorticoids, dexamethasone, prednisone, prednisolone, methylpred, betamethasone), carboxyamidotriazole, combretastatin A-4, squalamine, 6-O-chloroacetyl-carbonyl)-fumagillol, thalidomide, angiostatin, troponin-1, angiotensin II antagonists (see Fernandez et al.,  J. Lab. Clin. Med.  105:141-145 (1985)), and antibodies to VEGF (see,  Nature Biotechnology , Vol. 17, pp. 963-968 (October 1999); Kim et al.,  Nature,  362, 841-844 (1993); WO 00/44777; and WO 00/61186). 
         [0115]    Other examples of angiogenesis inhibitors include, but are not limited to, endostatin, ukrain, ranpirnase, IM862, 5-methoxy-4-[2-methyl-3-(3-methyl-2-butenyl)oxiranyl]-1-oxaspiro[2,5]oct-6-yl(chloroacetyl)carbamate, acetyldinanaline, 5-amino-1-[[3,5-dichloro-4-(4-chlorobenzoyl)phenyl]methyl]-1H-1,2,3-triazole-4-carboxamide, CM101, squalamine, combretastatin, RPI4610, NX31838, sulfated mannopentaose phosphate, 7,7-(carbonyl-bis[imino-N-methyl-4,2-pyrrolocarbonylimino[N-methyl-4,2-pyrrole]-carbonylimino]-bis-(1,3-naphthalene disulfonate), and 3-[(2,4-dimethylpyrrol-5-yl)methylene]-2-indolinone (SU5416). 
         [0116]    Other therapeutic agents that modulate or inhibit angiogenesis and may also be used in combination with WEE1 inhibitors include agents that modulate or inhibit the coagulation and fibrinolysis systems (see review in  Clin. Chem. La. Med.  38:679-692 (2000)). Examples of such agents that modulate or inhibit the coagulation and fibrinolysis pathways include, but are not limited to, heparin (see  Thromb. Haemost.  80:10-23 (1998)), low molecular weight heparins and carboxypeptidase U inhibitors (also known as inhibitors of active thrombin activatable fibrinolysis inhibitor [TAFIa]) (see  Thrombosis Res.  101:329-354 (2001)). TAFIa inhibitors have been described in U.S. Ser. Nos. 60/310,927 (filed Aug. 8, 2001) and 60/349,925 (filed Jan. 18, 2002). 
         [0117]    “Agents that interfere with cell cycle checkpoints” refer to compounds that inhibit protein kinases that transduce cell cycle checkpoint signals, thereby sensitizing the cancer cell to DNA damaging agents. Such agents include inhibitors of ATR, ATM, the CHK11 and CHK12 kinases and cdk and cdc kinase inhibitors and are specifically exemplified by 7-hydroxystaurosporin, flavopiridol, CYC202 (Cyclacel) and BMS-387032. 
         [0118]    “Agents that interfere with receptor tyrosine kinases (RTKs)” refer to compounds that inhibit RTKs and therefore mechanisms involved in oncogenesis and tumor progression. Such agents include inhibitors of c-Kit, Eph, PDGF, Flt3 and c-Met. Further agents include inhibitors of RTKs as described by Bume-Jensen and Hunter,  Nature,  411:355-365, 2001. 
         [0119]    “Inhibitors of cell proliferation and survival signalling pathway” refer to compounds that inhibit signal transduction cascades downstream of cell surface receptors. Such agents include inhibitors of serine/threonine kinases (including but not limited to inhibitors of Akt such as described in WO 02/083064, WO 02/083139, WO 02/083140, US 2004-0116432, WO 02/083138, US 2004-0102360, WO 03/086404, WO 03/086279, WO 03/086394, WO 03/084473, WO 03/086403, WO 2004/041162, WO 2004/096131, WO 2004/096129, WO 2004/096135, WO 2004/096130, WO 2005/100356, WO 2005/100344, US 2005/029941, US 2005/44294, US 2005/43361, 60/734,188, 60/652,737, 60/670,469), inhibitors of Raf kinase (for example PLX-4032), inhibitors of MEK (for example Arry-162, RO-4987655 and GSK-1120212), inhibitors of mTOR (for example AZD-8055, BEZ-235 and everolimus), and inhibitors of PI3K (for example GDC-0941, BKM-120). 
         [0120]    As used above, “integrin blockers” refers to compounds which selectively antagonize, inhibit or counteract binding of a physiological ligand to the α v β 3  integrin, to compounds which selectively antagonize, inhibit or counteract binding of a physiological ligand to the αvβ5 integrin, to compounds which antagonize, inhibit or counteract binding of a physiological ligand to both the α v β 3  integrin and the α v β 5  integrin, and to compounds which antagonize, inhibit or counteract the activity of the particular integrin(s) expressed on capillary endothelial cells. The term also refers to antagonists of the α v β 6 , α v β 8 , α 1 β 1 , α 2 β 1 , α 5 β 1 , α 6 β 1  and α 6 β 4  integrins. 
         [0121]    Some specific examples of tyrosine kinase inhibitors include N-(trifluoromethylphenyl)-5-methylisoxazol-4-carboxamide, 3-[(2,4-dimethylpyrrol-5-yl)methylidenyl)indolin-2-one, 17-(allylamino)-17-demethoxygeldanamycin, 4-(3-chloro-4-fluorophenylamino)-7-methoxy-6-[3-(4-morpholinyl)propoxyl]quinazoline, N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)-4-quinazolinamine, BIBX1382, 2,3,9,10,11,12-hexahydro-10-(hydroxymethyl)-10-hydroxy-9-methyl-9,12-epoxy-1H-diindolo[1,2,3-fg:3′,2′,1′-kl]pyrrolo[3,4-i][1,6]benzodiazocin-1-one, SH268, genistein, STI571, CEP2563, 4-(3-chlorophenylamino)-5,6-dimethyl-7H-pyrrolo[2,3-d]pyrimidinemethane sulfonate, 4-(3-bromo-4-hydroxyphenyl)amino-6,7-dimethoxyquinazoline, 4-(4′-hydroxyphenyl)amino-6,7-dimethoxyquinazoline, SU6668, STI571A, N-4-chlorophenyl-4-(4-pyridylmethyl)-1-phthalazinamine, and EMD121974. 
         [0122]    Combinations of the WEE1 inhibitor with PPAR-γ (i.e., PPAR-gamma) agonists and PPAR-δ (i.e., PPAR-delta) agonists may be useful in the treatment of certain malignancies. PPAR-γ and PPAR-δ are the nuclear peroxisome proliferator-activated receptors γ and δ. The expression of PPAR-γ on endothelial cells and its involvement in angiogenesis has been reported in the literature (see  J. Cardiovasc. Pharmacol.  1998; 31:909-913 ; J. Biol. Chem.  1999; 274:9116-9121 ; Invest. Ophthalmol Vis. Sci.  2000; 41:2309-2317). More recently, PPAR-γ agonists have been shown to inhibit the angiogenic response to VEGF in vitro; both troglitazone and rosiglitazone maleate inhibit the development of retinal neovascularization in mice. ( Arch. Ophthamol.  2001; 119:709-717). Examples of PPAR-γ agonists and PPAR-γ/α agonists include, but are not limited to, thiazolidinediones (such as DRF2725, CS-011, troglitazone, rosiglitazone, and pioglitazone), fenofibrate, gemfibrozil, clofibrate, GW2570, SB219994, AR-H039242, JTT-501, MCC-555, GW2331, GW409544, NN2344, KRP297, NP0110, DRF4158, NN622, GI262570, PNU182716, DRF552926, 2-[(5,7-dipropyl-3-trifluoromethyl-1,2-benzisoxazol-6-yl)oxy]-2-methylpropionic acid, and 2(R)-7-(3-(2-chloro-4-(4-fluorophenoxy) phenoxy)propoxy)-2-ethylchromane-2-carboxylic acid. 
         [0123]    Another embodiment of the instant invention is the use of WEE1 inhibitors in combination with gene therapy for the potential treatment of cancer. For an overview of genetic strategies to treating cancer see Hall et al ( Am. J. Hum. Genet.  61:785-789, 1997) and Kufe et al (Cancer Medicine, 5th Ed, pp 876-889, BC Decker, Hamilton 2000). Gene therapy can be used to deliver any tumor suppressing gene. Examples of such genes include, but are not limited to, p53, which can be delivered via recombinant virus-mediated gene transfer (see U.S. Pat. No. 6,069,134, for example), a uPA/uPAR antagonist (“Adenovirus-Mediated Delivery of a uPA/uPAR Antagonist Suppresses Angiogenesis-Dependent Tumor Growth and Dissemination in Mice,” Gene Therapy, August 1998; 5(8):1105-13), and interferon gamma ( J. Immunol.  2000; 164:217-222). 
         [0124]    The invention disclosed herein is exemplified by the following examples which should not be construed to limit the scope of the disclosure. 
       EXAMPLES 
     Example 1 
       [0125]    Thirty-one cell lines were treated with various DNA-damaging agents with or without WEE1-1 in a cell growth assay. Immediately after cells were plated, each drug was added at four concentrations for a total of 16 treatment conditions per combination (4×4 dose grid). Ninety-six hours following treatment, cell growth was evaluated with CellTiter-Glo (Promega) in treated samples relative to vehicle (DMSO) treated samples. Synergy was quantitated as the predicted additive growth inhibition of the two drugs (using the Bliss additivity model) subtracted from the observed growth inhibition of the two drugs. Therefore, a larger positive net difference indicates greater synergy, a negative difference indicates antagonism, and values at or close to 0 indicate additivity predicted by the Bliss model. The scale is displayed in the upper left corner of  FIG. 1 ; darkest grey denotes strong synergy, light grey denotes moderate synergy, and darker grey denotes values that are considered additive, not synergistic. Each column represents a different cell line; each row represents a pairing between WEE1-1 and different DNA-damaging agents. The cell line name is shown above the column and the tissue type from which the cell line was derived is at the bottom of the column. The 12 leftmost columns represent cell lines that are reported to be TP53 wildtype; the 19 rightmost columns represent cell lines that are reported to have mutations in TP53. Data are summarized in the table to the right. The p53+/+ column summarizes the percentage of p53+/+ cell lines that exhibited synergy in the assay (i.e., the number of darkest grey and light grey squares in a row divided by 12). The p53−/− column summarizes the percentage of p53−/− cell lines that exhibited synergy in the assay (i.e., the number of darkest grey and light grey squares in a row divided by 19). Overall, the percentage of p53 wildtype cell lines that exhibited synergy was 15/(12×9)=14%; the percentage of P53 mutant cell lines that exhibited synergy was 50/(19×9)=29%. Thus, in combination with DNA-damaging agents, WEE1-1 is more likely to lead to synergistic growth inhibition in cell lines defective in TP53. 
       Example 2 
     Identification of Loss of Function Mutations 
       [0126]    A multiple-step approach to develop a list of mutations which would be most likely to result in a non-functional p53 protein was implemented. First, different types of mutations predicted to result in p53 loss-of-function were assigned a relative point value, as shown in the Table 1 below. For example, three points were assigned for any mutation that results in a truncation, frameshift or a splice site defect that would completely eliminate the DNA binding domain of p53, i.e., before amino acid 306. (A frameshift mutation is one that causes the ribosome to use a different reading frame on the mRNA. For example, frameshifts can occur from one or two basepair insertions or deletions). Second, published data in the International Agency for Research on Cancer (IARC) database (A. Petijean et al.  Hum Mutat.  2007, 28(6):622-9) was used to assess the impact of each possible mutation on p53 function (dominant negative protein or transactivation function). Mutations shown to display dominant negative activity in a model system were assigned a point value of 2.0 points, while mutations reducing or eliminating the protein&#39;s transactivation function were assigned a value of 1 point. Third, data from an internal gene expression database and the Cell line Biomarker Discovery (CBD) expression database of 650 cancer cell lines was used to identify mutations in TP53 that resulted in gene expression patterns indicative of p53 loss-of-function. These mutations were assigned 1 point. Finally, mutational analysis of tumor samples reported to the IARC database was used to identify mutations that were observed in more than 10 somatic (i.e., tumor) samples, which were assigned a point value of 0.5 points. 
         [0000]    
       
         
               
               
             
               
               
             
           
               
                 TABLE 1 
               
               
                   
               
               
                 Type of Mutation 
                 Point Value 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 Stop codon mutation (truncated protein) prior to codon 306 
                 3 
               
               
                 Splice site mutation between codons 97and 375 
                 3 
               
               
                 Frameshift mutation 
                 3 
               
               
                 Dominant negative mutation 
                 2 
               
               
                 P53 signature score p-value &lt; 0.05 vs WT 
                 1 
               
               
                 Transactivation loss-of-function mutation 
                 1 
               
               
                 Mutation reported &gt;10 times in somatic tissue at 
                 0.5 
               
               
                 nucleotide level 
               
               
                 Mutation reported &gt;10 times in somatic tissue at 
                 0.5 
               
               
                 amino acid level 
               
               
                   
               
             
          
         
       
     
         [0000]    
       
         
               
             
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 2 
               
               
                   
               
               
                 Mutation Lookup Table 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                 E2* 
                 K120* 
                 V143M 
                 A159D 
                 C176F 
                 R196P 
                 C229* 
                 C242S 
                 R249S 
                 C275G 
                 R282G 
               
               
                 E3* 
                 C124* 
                 V143A 
                 A161D 
                 C176Y 
                 V197G 
                 Y234H 
                 C242* 
                 P250L 
                 C275Y 
                 R282W 
               
               
                 Q5* 
                 Y126* 
                 Q144* 
                 Y163N 
                 C176S 
                 E198* 
                 Y234C 
                 C242W 
                 I255F 
                 C275F 
                 R282P 
               
               
                 S6* 
                 S127P 
                 L145Q 
                 Y163H 
                 C176* 
                 G199* 
                 Y234* 
                 G244S 
                 L257Q 
                 C275S 
                 R282Q 
               
               
                 E11* 
                 S127T 
                 L145P 
                 Y163D 
                 C176W 
                 G199E 
                 Y236D 
                 G244C 
                 L257P 
                 C275* 
                 R283H 
               
               
                 Q16* 
                 S127F 
                 W146* 
                 Y163C 
                 P177S 
                 L201* 
                 Y236H 
                 G244R 
                 E258* 
                 C275W 
                 R283P 
               
               
                 E17* 
                 S127Y 
                 D148* 
                 Y163S 
                 P177H 
                 E204* 
                 Y236N 
                 G244D 
                 E258K 
                 A276P 
                 E285* 
               
               
                 S20* 
                 S127C 
                 P151T 
                 Y163* 
                 P177R 
                 Y205D 
                 Y236C 
                 G244V 
                 D259Y 
                 C277Y 
                 E285K 
               
               
                 W23* 
                 K132* 
                 P151S 
                 K164* 
                 H179N 
                 Y205S 
                 Y236S 
                 G244A 
                 D259V 
                 C277F 
                 E285V 
               
               
                 K24* 
                 K132Q 
                 P151A 
                 K164E 
                 H179Y 
                 Y205C 
                 Y236* 
                 G245S 
                 N263* 
                 C277* 
                 E286* 
               
               
                 E28* 
                 K132E 
                 P151R 
                 Q165* 
                 H179R 
                 Y205* 
                 M237I 
                 G245R 
                 L265P 
                 P278T 
                 E286K 
               
               
                 L35* 
                 K132R 
                 P151H 
                 S166* 
                 H179L 
                 L206* 
                 C238R 
                 G245C 
                 G266* 
                 P278S 
                 E286Q 
               
               
                 Q38* 
                 K132M 
                 P152S 
                 Q167* 
                 H179Q 
                 R209* 
                 C238Y 
                 G245D 
                 G266R 
                 P278A 
                 E286G 
               
               
                 L43* 
                 K132T 
                 P152L 
                 H168Y 
                 H179* 
                 R213* 
                 C238F 
                 G245V 
                 G266E 
                 P278H 
                 E286V 
               
               
                 E51* 
                 K132N 
                 G154V 
                 H168R 
                 E180* 
                 R213Q 
                 C238* 
                 G245A 
                 G266V 
                 P278L 
                 E286A 
               
               
                 Q52* 
                 M133K 
                 T155P 
                 H168P 
                 C182* 
                 R213L 
                 N239D 
                 M246V 
                 R267W 
                 P278R 
                 E286D 
               
               
                 W53* 
                 C135R 
                 T155N 
                 H168L 
                 S183* 
                 H214R 
                 N239S 
                 M246L 
                 R267P 
                 G279E 
                 E287* 
               
               
                 E56* 
                 C135S 
                 R156P 
                 E171* 
                 S183* 
                 S215I 
                 N239* 
                 M246R 
                 F270C 
                 R280* 
                 N288D 
               
               
                 E62* 
                 C135G 
                 V157F 
                 V173M 
                 Q192* 
                 V216M 
                 S240G 
                 N247I 
                 E271* 
                 R280G 
                 K291* 
               
               
                 R65* 
                 C135Y 
                 V157L 
                 V173L 
                 H193Y 
                 V216L 
                 S241T 
                 N247T 
                 V272M 
                 R280T 
                 K292* 
               
               
                 E68* 
                 C135F 
                 V157D 
                 V173G 
                 H193D 
                 V216E 
                 S241A 
                 R248W 
                 R273C 
                 R280K 
                 E294* 
               
               
                 W91* 
                 C135S 
                 V157G 
                 V173E 
                 H193N 
                 V216G 
                 S241P 
                 R248G 
                 R273S 
                 R280I 
                 E298* 
               
               
                 S94* 
                 C135* 
                 V157A 
                 R175G 
                 H193L 
                 Y220N 
                 S241F 
                 R248Q 
                 R273G 
                 R280S 
                 K305* 
               
               
                 Q100* 
                 C135W 
                 R158G 
                 R175C 
                 H193P 
                 Y220H 
                 S241C 
                 R248L 
                 R273H 
                 D281N 
                 R306* 
               
               
                 K101* 
                 Q136* 
                 R158S 
                 R175H 
                 H193R 
                 Y220D 
                 S241Y 
                 R248P 
                 R273L 
                 D281Y 
                 R337C 
               
               
                 Y103* 
                 A138P 
                 R158P 
                 R175L 
                 L194F 
                 Y220C 
                 C242S 
                 R249W 
                 R273P 
                 D281H 
               
               
                 Q104* 
                 A138G 
                 R158H 
                 R175P 
                 L194R 
                 Y220S 
                 C242R 
                 R249G 
                 V274F 
                 D281G 
               
               
                 Y107* 
                 K139* 
                 R158L 
                 C176R 
                 I195F 
                 Y220* 
                 C242G 
                 R249M 
                 V274A 
                 D281V 
               
               
                 R110L 
                 C141Y 
                 A159P 
                 C176S 
                 I195T 
                 E221* 
                 C242Y 
                 R249K 
                 C275R 
                 D281A 
               
               
                 L114* 
                 C141* 
                 A159V 
                 C176G 
                 R196* 
                 E224* 
                 C242F 
                 R249T 
                 C275S 
                 D281E 
               
               
                   
               
               
                 PLUS ANY MUTATIONS LABELED “FRAMESHIFT” OR “SPLICE DEFECT” 
               
               
                 *Denotes stop codon; The GENBANK accession number of the p53 protein is NM000546. The GENBANK accession number of the TP53 gene is X54156.1 
               
             
          
         
       
     
         [0127]    The TP53 mutations in the look-up table (Table 2) include those whose evidence score is 2.0 or greater. The expanded table of TP53 mutations (Table 3) shows the location and type of mutation, as well as the point value assigned to the mutation. 
         [0000]    
       
         
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
               
             
               
               
               
               
               
               
               
               
               
               
               
               
               
             
           
               
                 TABLE 3 
               
               
                   
               
               
                 Supporting evidence for mutations in the Mutation Lookup Table 
               
               
                   
               
             
             
               
                   
               
             
          
           
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 P53 
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 signa- 
                 Reported &gt;10x 
                 Reported &gt;10x 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 Trans- 
                 P53 
                 ture 
                 in somatic 
                 in somatic 
               
               
                   
                   
                 WT 
                   
                 Pro- 
                 Stop 
                 Splice 
                 Dominant 
                 activa- 
                 Signa- 
                 p value &lt; 
                 @ AA 
                 @ Base 
                 Evi- 
               
               
                   
                 Co- 
                 Co- 
                 Mutant 
                 tein 
                 Codon 
                 Defect 
                 Negative 
                 tion 
                 ture 
                 0.05 
                 level 
                 level 
                 dence 
               
               
                 NtNum 
                 don 
                 don 
                 Codon 
                 Change 
                 (3 pts) 
                 (3 pts) 
                 (2 pts) 
                 (1 pt) 
                 P-Value 
                 (1 pt) 
                 (0.5 pt) 
                 (0.5 pt) 
                 Score 
               
               
                   
               
               
                 11009 
                 2 
                 GAG 
                 TAG 
                 E2* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 0 
                 3 
               
               
                 11012 
                 3 
                 GAG 
                 TAG 
                 E3* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 0 
                 3 
               
               
                 11018 
                 5 
                 CAG 
                 TAG 
                 Q5* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 1 
                 3 
               
               
                 11022 
                 6 
                 TCA 
                 TAA 
                 S6* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 0 
                 3 
               
               
                 11022 
                 6 
                 TCA 
                 TGA 
                 S6* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 0 
                 3 
               
               
                 11036 
                 11 
                 GAG 
                 TAG 
                 E11* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 0 
                 3 
               
               
                 11051 
                 16 
                 CAG 
                 TAG 
                 Q16* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 0 
                 3 
               
               
                 11054 
                 17 
                 GAA 
                 TAA 
                 E17* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 0 
                 3 
               
               
                 11064 
                 20 
                 TCA 
                 TAA 
                 S20* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 0 
                 3 
               
               
                 11064 
                 20 
                 TCA 
                 TGA 
                 S20* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 0 
                 3 
               
               
                 11073 
                 23 
                 TGG 
                 TAG 
                 W23* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 0 
                 3 
               
               
                 11074 
                 23 
                 TGG 
                 TGA 
                 W23* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 0 
                 3 
               
               
                 11075 
                 24 
                 AAA 
                 TAA 
                 K24* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 0 
                 3 
               
               
                 11204 
                 28 
                 GAA 
                 TAA 
                 E28* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 0 
                 3 
               
               
                 11335 
                 35 
                 TTG 
                 TAG 
                 L35* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 0 
                 3 
               
               
                 11343 
                 38 
                 CAA 
                 TAA 
                 Q38* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
                 3 
               
               
                 11359 
                 43 
                 TTG 
                 TAG 
                 L43* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
                 3 
               
               
                 11382 
                 51 
                 GAA 
                 TAA 
                 E51* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 6 
                 3 
               
               
                 11385 
                 52 
                 CAA 
                 TAA 
                 Q52* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 8 
                 3 
               
               
                 11389 
                 53 
                 TGG 
                 TAG 
                 W53* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 7 
                 3 
               
               
                 11390 
                 53 
                 TGG 
                 TGA 
                 W53* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 10 
                 3 
               
               
                 11397 
                 56 
                 GAA 
                 TAA 
                 E56* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 6 
                 3 
               
               
                 11415 
                 62 
                 GAA 
                 TAA 
                 E62* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 9 
                 3 
               
               
                 11424 
                 65 
                 AGA 
                 TGA 
                 R65* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
                 3 
               
               
                 11433 
                 68 
                 GAG 
                 TAG 
                 E68* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 6 
                 3 
               
               
                 11503 
                 91 
                 TGG 
                 TAG 
                 W91* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 13 
                 3.5 
               
               
                 11504 
                 91 
                 TGG 
                 TGA 
                 W91* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 12 
                 3.5 
               
               
                 11512 
                 94 
                 TCA 
                 TAA 
                 S94* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 2 
                 3 
               
               
                 11512 
                 94 
                 TCA 
                 TGA 
                 S94* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 2 
                 3 
               
               
                 11529 
                 100 
                 CAG 
                 TAG 
                 Q100* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 16 
                 3.5 
               
               
                 11532 
                 101 
                 AAA 
                 TAA 
                 K101* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 1 
                 3 
               
               
                 11540 
                 103 
                 TAC 
                 TAA 
                 Y103* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
                 3 
               
               
                 11540 
                 103 
                 TAC 
                 TAG 
                 Y103* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 1 
                 3 
               
               
                 11541 
                 104 
                 CAG 
                 TAG 
                 Q104* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 16 
                 3.5 
               
               
                 11552 
                 107 
                 TAC 
                 TAA 
                 Y107* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 4 
                 3 
               
               
                 11552 
                 107 
                 TAC 
                 TAG 
                 Y107* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 5 
                 3 
               
               
                 11560 
                 110 
                 CGT 
                 CTT 
                 R110L 
                   
                   
                   
                 Non 
                 1.00E−01 
                   
                 18 
                 30 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 11572 
                 114 
                 TTG 
                 TAG 
                 L114* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
                 3 
               
               
                 11589 
                 120 
                 AAG 
                 TAG 
                 K120* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 2 
                 3 
               
               
                 11603 
                 124 
                 TGC 
                 TGA 
                 C124* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 1 
                 3 
               
               
                 12366 
                 126 
                 TAC 
                 TAA 
                 Y126* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 12 
                 3.5 
               
               
                 12366 
                 126 
                 TAC 
                 TAG 
                 Y126* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 13 
                 3.5 
               
               
                 12367 
                 127 
                 TCC 
                 CCC 
                 S127P 
                   
                   
                 Yes 
                 Non 
                 1.00E−02 
                 Yes 
                 3 
                 6 
                 4 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12367 
                 127 
                 TCC 
                 ACC 
                 S127T 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                   
                 3 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12368 
                 127 
                 TCC 
                 TTC 
                 S127F 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 2 
                 23 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12368 
                 127 
                 TCC 
                 TAC 
                 S127Y 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 5 
                 9 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12368 
                 127 
                 TCC 
                 TGC 
                 S127C 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 1 
                 2 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12382 
                 132 
                 AAG 
                 TAG 
                 K132* 
                 Yes 
                   
                   
                   
                 7.00E−04 
                 Yes 
                   
                 2 
                 4 
               
               
                 12382 
                 132 
                 AAG 
                 CAG 
                 K132Q 
                   
                   
                   
                 Non 
                 7.00E−04 
                 Yes 
                 12 
                 17 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12382 
                 132 
                 AAG 
                 GAG 
                 K132E 
                   
                   
                   
                 Non 
                 7.00E−04 
                 Yes 
                 16 
                 24 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12383 
                 132 
                 AAG 
                 AGG 
                 K132R 
                   
                   
                 Yes 
                 Non 
                 7.00E−04 
                 Yes 
                 30 
                 57 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12383 
                 132 
                 AAG 
                 ATG 
                 K132M 
                   
                   
                   
                 Non 
                 7.00E−04 
                 Yes 
                 7 
                 12 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12383 
                 132 
                 AAG 
                 ACG 
                 K132T 
                   
                   
                   
                 Non 
                 7.00E−04 
                 Yes 
                 3 
                 4 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12384 
                 132 
                 AAG 
                 AAT 
                 K132N 
                   
                   
                 Yes 
                 Non 
                 7.00E−04 
                 Yes 
                 22 
                 26 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12384 
                 132 
                 AAG 
                 AAC 
                 K132N 
                   
                   
                 Yes 
                 Non 
                 7.00E−04 
                 Yes 
                 22 
                 34 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12386 
                 133 
                 ATG 
                 AAG 
                 M133K 
                   
                   
                   
                 Non 
                 2.00E−01 
                   
                 11 
                 20 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12391 
                 135 
                 TGC 
                 CGC 
                 C135R 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 5 
                 16 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12391 
                 135 
                 TGC 
                 AGC 
                 C135S 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 4 
                 8 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12391 
                 135 
                 TGC 
                 GGC 
                 C135G 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 6 
                 9 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12392 
                 135 
                 TGC 
                 TAC 
                 C135Y 
                   
                   
                 Yes 
                 Non 
                 2.00E−02 
                 Yes 
                 46 
                 80 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12392 
                 135 
                 TGC 
                 TTC 
                 C135F 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 34 
                 56 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12392 
                 135 
                 TGC 
                 TCC 
                 C135S 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 4 
                 8 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12393 
                 135 
                 TGC 
                 TGA 
                 C135* 
                 Yes 
                   
                   
                   
                 2.00E−02 
                 Yes 
                   
                 8 
                 4 
               
               
                 12393 
                 135 
                 TGC 
                 TGG 
                 C135W 
                   
                   
                   
                   
                 2.00E−02 
                 Yes 
                 17 
                 26 
                 2 
               
               
                 12394 
                 136 
                 CAA 
                 TAA 
                 Q136* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 39 
                 3.5 
               
               
                 12400 
                 138 
                 GCC 
                 CCC 
                 A138P 
                   
                   
                   
                 Non 
                 4.00E−02 
                 Yes 
                 12 
                 30 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12401 
                 138 
                 GCC 
                 GGC 
                 A138G 
                   
                   
                   
                 Non 
                 4.00E−02 
                 Yes 
                   
                 0 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12403 
                 139 
                 AAG 
                 TAG 
                 K139* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 5 
                 3 
               
               
                 12410 
                 141 
                 TGC 
                 TAC 
                 C141Y 
                   
                   
                   
                 Non 
                 5.00E−01 
                   
                 61 
                 97 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12411 
                 141 
                 TGC 
                 TGA 
                 C141* 
                 Yes 
                   
                   
                   
                 5.00E−01 
                   
                   
                 19 
                 3.5 
               
               
                 12415 
                 143 
                 GTG 
                 ATG 
                 V143M 
                   
                   
                   
                 Non 
                 1.00E−01 
                   
                 15 
                 30 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12416 
                 143 
                 GTG 
                 GCG 
                 V143A 
                   
                   
                   
                 Non 
                 1.00E−01 
                   
                 13 
                 20 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12418 
                 144 
                 CAG 
                 TAG 
                 Q144* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 44 
                 3.5 
               
               
                 12422 
                 145 
                 CTG 
                 CAG 
                 L145Q 
                   
                   
                   
                 Non 
                   
                   
                 14 
                 19 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12422 
                 145 
                 CTG 
                 CCG 
                 L145P 
                   
                   
                   
                 Non 
                   
                   
                 15 
                 21 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12425 
                 146 
                 TGG 
                 TAG 
                 W146* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 50 
                 3.5 
               
               
                 12426 
                 146 
                 TGG 
                 TGA 
                 W146* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 51 
                 3.5 
               
               
                 12432 
                 148 
                 GAT 
                 TGA 
                 D148* 
                 Yes 
                   
                   
                 Non 
                 6.00E−03 
                 Yes 
                   
                   
                 4 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12439 
                 151 
                 CCC 
                 ACC 
                 P151T 
                   
                   
                   
                 Non 
                 6.00E−03 
                 Yes 
                 13 
                 21 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12439 
                 151 
                 CCC 
                 TCC 
                 P151S 
                   
                   
                   
                 Non 
                 6.00E−03 
                 Yes 
                 58 
                 95 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12439 
                 151 
                 CCC 
                 GCC 
                 P151A 
                   
                   
                   
                 Non 
                 6.00E−03 
                 Yes 
                 7 
                 18 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12440 
                 151 
                 CCC 
                 CGC 
                 P151R 
                   
                   
                 Yes 
                 Non 
                 6.00E−03 
                 Yes 
                 6 
                 20 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12440 
                 151 
                 CCC 
                 CAC 
                 P151H 
                   
                   
                   
                 Non 
                 6.00E−03 
                 Yes 
                 1 
                 36 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12442 
                 152 
                 CCG 
                 TCG 
                 P152S 
                   
                   
                   
                 Non 
                 6.00E−01 
                   
                 21 
                 30 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12442 
                 152 
                 CCG 
                 CTG 
                 P152L 
                   
                   
                   
                 Non 
                 6.00E−03 
                 Yes 
                 84 
                 10 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12449 
                 154 
                 GGC 
                 GTC 
                 G154V 
                   
                   
                   
                 Non 
                 4.00E−01 
                   
                 34 
                 64 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12451 
                 155 
                 ACC 
                 CCC 
                 T155P 
                   
                   
                   
                 Non 
                 1.00E−01 
                   
                 13 
                 21 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12452 
                 155 
                 ACC 
                 AAC 
                 T155N 
                   
                   
                   
                 Non 
                 1.00E−01 
                   
                 19 
                 32 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12455 
                 156 
                 CGC 
                 CCC 
                 R156P 
                   
                   
                   
                 Non 
                 8.00E−01 
                   
                 23 
                 44 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12457 
                 157 
                 GTC 
                 TTC 
                 V157F 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 133 
                 186 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12457 
                 157 
                 GTC 
                 CTC 
                 V157L 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 6 
                 9 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12458 
                 157 
                 GTC 
                 GAC 
                 V157D 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 8 
                 14 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12458 
                 157 
                 GTC 
                 GGC 
                 V157G 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 7 
                 12 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12458 
                 157 
                 GTC 
                 GCC 
                 V157A 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 1 
                 2 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12460 
                 158 
                 CGC 
                 GGC 
                 R158G 
                   
                   
                   
                 Non 
                 4.00E−03 
                 Yes 
                 10 
                 22 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12460 
                 158 
                 CGC 
                 AGC 
                 R158S 
                   
                   
                   
                 Non 
                 4.00E−03 
                 Yes 
                   
                 4 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12461 
                 158 
                 CGC 
                 CCC 
                 R158P 
                   
                   
                 Yes 
                 Non 
                 4.00E−03 
                 Yes 
                 9 
                 19 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12461 
                 158 
                 CGC 
                 CAC 
                 R158H 
                   
                   
                   
                 Non 
                 4.00E−03 
                 Yes 
                 55 
                 106 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12461 
                 158 
                 CGC 
                 CTC 
                 R158L 
                   
                   
                   
                 Non 
                 4.00E−03 
                 Yes 
                 1 
                 96 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12463 
                 159 
                 GCC 
                 CCC 
                 A159P 
                   
                   
                   
                 Non 
                 3.00E−03 
                 Yes 
                 13 
                 30 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12464 
                 159 
                 GCC 
                 GTC 
                 A159V 
                   
                   
                   
                 Non 
                 3.00E−03 
                 Yes 
                 28 
                 46 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12464 
                 159 
                 GCC 
                 GAC 
                 A159D 
                   
                   
                   
                 Non 
                 3.00E−03 
                 Yes 
                 6 
                 11 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12470 
                 161 
                 GCC 
                 GAC 
                 A161D 
                   
                   
                   
                 Non 
                 4.00E−02 
                 Yes 
                 7 
                 21 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12475 
                 163 
                 TAC 
                 AAC 
                 Y163N 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 16 
                 23 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12475 
                 163 
                 TAC 
                 CAC 
                 Y163H 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 16 
                 25 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12475 
                 163 
                 TAC 
                 GAC 
                 Y163D 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 2 
                 4 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12476 
                 163 
                 TAC 
                 TGC 
                 Y163C 
                   
                   
                 Yes 
                 Non 
                 2.00E−02 
                 Yes 
                 90 
                 147 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12476 
                 163 
                 TAC 
                 TCC 
                 Y163S 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 4 
                 5 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12477 
                 163 
                 TAC 
                 TAG 
                 Y163* 
                 Yes 
                   
                   
                   
                 2.00E−02 
                 Yes 
                   
                 11 
                 4.5 
               
               
                 12477 
                 163 
                 TAC 
                 TAA 
                 Y163* 
                 Yes 
                   
                   
                   
                 2.00E−02 
                 Yes 
                   
                 7 
                 4 
               
               
                 12478 
                 164 
                 AAG 
                 TAG 
                 K164* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 17 
                 3.5 
               
               
                 12478 
                 164 
                 AAG 
                 GAG 
                 K164E 
                   
                   
                   
                 Non 
                   
                   
                 11 
                 24 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12481 
                 165 
                 CAG 
                 TAG 
                 Q165* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 45 
                 3.5 
               
               
                 12485 
                 166 
                 TCA 
                 TAA 
                 S166* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 15 
                 3.5 
               
               
                 12485 
                 166 
                 TCA 
                 TGA 
                 S166* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 15 
                 3.5 
               
               
                 12487 
                 167 
                 CAG 
                 TAG 
                 Q167* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 38 
                 3.5 
               
               
                 12490 
                 168 
                 CAC 
                 TAC 
                 H168Y 
                   
                   
                 Yes 
                   
                 1.00E−02 
                 Yes 
                 9 
                 14 
                 3.5 
               
               
                 12491 
                 168 
                 CAC 
                 CGC 
                 H168R 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 12 
                 22 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12491 
                 168 
                 CAC 
                 CCC 
                 H168P 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 9 
                 13 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12491 
                 168 
                 CAC 
                 CTC 
                 H168L 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 5 
                 8 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12499 
                 171 
                 GAG 
                 TAG 
                 E171* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 21 
                 3.5 
               
               
                 12505 
                 173 
                 GTG 
                 ATG 
                 V173M 
                   
                   
                 Yes 
                 Non 
                 1.00E−02 
                 Yes 
                 35 
                 71 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12505 
                 173 
                 GTG 
                 TTG 
                 V173L 
                   
                   
                 Yes 
                 Non 
                 1.00E−02 
                 Yes 
                 9 
                 65 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12505 
                 173 
                 GTG 
                 CTG 
                 V173L 
                   
                   
                 Yes 
                 Non 
                 1.00E−02 
                 Yes 
                 9 
                 21 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12506 
                 173 
                 GTG 
                 GGG 
                 V173G 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 4 
                 16 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12506 
                 173 
                 GTG 
                 GAG 
                 V173E 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 1 
                 3 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12511 
                 175 
                 CGC 
                 GGC 
                 R175G 
                   
                   
                   
                 Non 
                 9.00E−13 
                 Yes 
                 11 
                 24 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12511 
                 175 
                 CGC 
                 TGC 
                 R175C 
                   
                   
                   
                   
                 9.00E−13 
                 Yes 
                 12 
                 27 
                 2 
               
               
                 12512 
                 175 
                 CGC 
                 CAC 
                 R175H 
                   
                   
                 Yes 
                 Non 
                 9.00E−13 
                 Yes 
                 691 
                 1158 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12512 
                 175 
                 CGC 
                 CTC 
                 R175L 
                   
                   
                   
                   
                 9.00E−13 
                 Yes 
                 18 
                 27 
                 2 
               
               
                 12512 
                 175 
                 CGC 
                 CCC 
                 R175P 
                   
                   
                   
                 Non 
                 9.00E−13 
                 Yes 
                 5 
                 8 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12514 
                 176 
                 TGC 
                 CGC 
                 C176R 
                   
                   
                 Yes 
                 Non 
                 2.00E−03 
                 Yes 
                 8 
                 14 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12514 
                 176 
                 TGC 
                 AGC 
                 C176S 
                   
                   
                   
                 Non 
                 2.00E−03 
                 Yes 
                 1 
                 20 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12514 
                 176 
                 TGC 
                 GGC 
                 C176G 
                   
                   
                   
                 Non 
                 2.00E−03 
                 Yes 
                 3 
                 7 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12515 
                 176 
                 TGC 
                 TTC 
                 C176F 
                   
                   
                 Yes 
                   
                 2.00E−03 
                 Yes 
                 1 
                 156 
                 3.5 
               
               
                 12515 
                 176 
                 TGC 
                 TAC 
                 C176Y 
                   
                   
                   
                 Non 
                 2.00E−03 
                 Yes 
                 44 
                 89 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12515 
                 176 
                 TGC 
                 TCC 
                 C176S 
                   
                   
                   
                 Non 
                 2.00E−03 
                 Yes 
                 1 
                 11 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12516 
                 176 
                 TGC 
                 TGA 
                 C176* 
                 Yes 
                   
                   
                   
                 2.00E−03 
                 Yes 
                   
                 9 
                 4 
               
               
                 12516 
                 176 
                 TGC 
                 TGG 
                 C176W 
                   
                   
                   
                 Non 
                 2.00E−03 
                 Yes 
                 11 
                 20 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12517 
                 177 
                 CCC 
                 TCC 
                 P177S 
                   
                   
                 Yes 
                   
                 7.00E−01 
                   
                 8 
                 16 
                 2.5 
               
               
                 12518 
                 177 
                 CCC 
                 CAC 
                 P177H 
                   
                   
                 Yes 
                   
                 7.00E−01 
                   
                 2 
                 5 
                 2 
               
               
                 12518 
                 177 
                 CCC 
                 CGC 
                 P177R 
                   
                   
                   
                 Non 
                 7.00E−01 
                   
                 13 
                 17 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12523 
                 179 
                 CAT 
                 AAT 
                 H179N 
                   
                   
                 Yes 
                   
                 2.00E−02 
                 Yes 
                 13 
                 24 
                 4 
               
               
                 12523 
                 179 
                 CAT 
                 TAT 
                 H179Y 
                   
                   
                 Yes 
                   
                 2.00E−02 
                 Yes 
                 8 
                 107 
                 3.5 
               
               
                 12524 
                 179 
                 CAT 
                 CGT 
                 H179R 
                   
                   
                 Yes 
                 Non 
                 2.00E−02 
                 Yes 
                 90 
                 151 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12524 
                 179 
                 CAT 
                 CTT 
                 H179L 
                   
                   
                 Yes 
                   
                 2.00E−02 
                 Yes 
                 31 
                 42 
                 4 
               
               
                 12525 
                 179 
                 CAT 
                 CAG 
                 H179Q 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 7 
                 16 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12525 
                 179 
                 CAT 
                 CAA 
                 H179Q 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 7 
                 9 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12525 
                 179 
                 CAT 
                 TAG 
                 H179* 
                 Yes 
                   
                   
                 Non 
                 7.00E−01 
                   
                 1 
                   
                 4 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12526 
                 180 
                 GAG 
                 TAG 
                 E180* 
                 Yes 
                   
                   
                   
                 2.00E−01 
                   
                   
                 19 
                 3.5 
               
               
                 12534 
                 182 
                 TGC 
                 TGA 
                 C182* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 7 
                 3 
               
               
                 12536 
                 183 
                 TCA 
                 TGA 
                 S183* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 26 
                 3.5 
               
               
                 12536 
                 183 
                 TCA 
                 TAA 
                 S183* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
                 3 
               
               
                 12643 
                 192 
                 CAG 
                 TAG 
                 Q192* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 96 
                 3.5 
               
               
                 12646 
                 193 
                 CAT 
                 TAT 
                 H193Y 
                   
                   
                   
                 Non 
                 5.00E−03 
                 Yes 
                 25 
                 39 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12646 
                 193 
                 CAT 
                 GAT 
                 H193D 
                   
                   
                   
                 Non 
                 5.00E−03 
                 Yes 
                 7 
                 15 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12646 
                 193 
                 CAT 
                 AAT 
                 H193N 
                   
                   
                   
                 Non 
                 5.00E−03 
                 Yes 
                 3 
                 3 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12647 
                 193 
                 CAT 
                 CTT 
                 H193L 
                   
                   
                   
                 Non 
                 5.00E−03 
                 Yes 
                 28 
                 60 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12647 
                 193 
                 CAT 
                 CCT 
                 H193P 
                   
                   
                   
                 Non 
                 5.00E−03 
                 Yes 
                 12 
                 18 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12647 
                 193 
                 CAT 
                 CGT 
                 H193R 
                   
                   
                   
                 Non 
                 5.00E−03 
                 Yes 
                 58 
                 91 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12649 
                 194 
                 CTT 
                 TTT 
                 L194F 
                   
                   
                   
                 Non 
                 2.00E−01 
                   
                 15 
                 27 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12650 
                 194 
                 CTT 
                 CGT 
                 L194R 
                   
                   
                   
                 Non 
                 2.00E−01 
                   
                 26 
                 60 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12652 
                 195 
                 ATC 
                 TTC 
                 I195F 
                   
                   
                   
                 Non 
                 3.00E−01 
                   
                 14 
                 28 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12653 
                 195 
                 ATC 
                 ACC 
                 I195T 
                   
                   
                   
                 Non 
                 3.00E−01 
                   
                 51 
                 89 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12655 
                 196 
                 CGA 
                 TGA 
                 R196* 
                 Yes 
                   
                   
                   
                 2.00E−01 
                   
                   
                 223 
                 3.5 
               
               
                 12656 
                 196 
                 CGA 
                 CCA 
                 R196P 
                   
                   
                   
                 Non 
                 2.00E−01 
                   
                 12 
                 19 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12659 
                 197 
                 GTG 
                 GGG 
                 V197G 
                   
                   
                   
                 Non 
                 2.00E−01 
                   
                 14 
                 22 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12661 
                 198 
                 GAA 
                 TAA 
                 E198* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 24 
                 3.5 
               
               
                 12664 
                 199 
                 GGA 
                 TGA 
                 G199* 
                 Yes 
                   
                   
                   
                 5.00E−01 
                   
                   
                 4 
                 3 
               
               
                 12665 
                 199 
                 GGA 
                 GAA 
                 G199E 
                   
                   
                   
                 Non 
                 5.00E−01 
                   
                 11 
                 15 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12671 
                 201 
                 TTG 
                 TAG 
                 L201* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 7 
                 3 
               
               
                 12679 
                 204 
                 GAG 
                 TAG 
                 E204* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 45 
                 3.5 
               
               
                 12682 
                 205 
                 TAT 
                 GAT 
                 Y205D 
                   
                   
                   
                 Non 
                 7.00E−02 
                   
                 13 
                 18 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12683 
                 205 
                 TAT 
                 TCT 
                 Y205S 
                   
                   
                   
                 Non 
                 7.00E−02 
                   
                 11 
                 20 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12683 
                 205 
                 TAT 
                 TGT 
                 Y205C 
                   
                   
                   
                 Non 
                 7.00E−02 
                   
                 48 
                 110 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12684 
                 205 
                 TAT 
                 TAA 
                 Y205* 
                 Yes 
                   
                   
                   
                 7.00E−02 
                   
                   
                 4 
                 3 
               
               
                 12684 
                 205 
                 TAT 
                 TAG 
                 Y205* 
                 Yes 
                   
                   
                   
                 7.00E−02 
                   
                   
                 4 
                 3 
               
               
                 12686 
                 206 
                 TTG 
                 TAG 
                 L206* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 11 
                 3.5 
               
               
                 12694 
                 209 
                 AGA 
                 TGA 
                 R209* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 13 
                 3.5 
               
               
                 12706 
                 213 
                 CGA 
                 TGA 
                 R213* 
                 Yes 
                   
                   
                   
                 6.00E−01 
                   
                   
                 5 
                 3 
               
               
                 12707 
                 213 
                 CGA 
                 CAA 
                 R213Q 
                   
                   
                   
                 Non 
                 6.00E−01 
                   
                 21 
                 36 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12707 
                 213 
                 CGA 
                 CTA 
                 R213L 
                   
                   
                   
                 Non 
                 6.00E−01 
                   
                 24 
                 38 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12710 
                 214 
                 CAT 
                 CGT 
                 H214R 
                   
                   
                   
                 Non 
                 2.00E−01 
                   
                 41 
                 78 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12713 
                 215 
                 AGT 
                 ATT 
                 S215I 
                   
                   
                   
                 Non 
                 9.00E−02 
                   
                 14 
                 25 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12715 
                 216 
                 GTG 
                 ATG 
                 V216M 
                   
                   
                   
                 Non 
                 7.00E−04 
                 Yes 
                 43 
                 76 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12715 
                 216 
                 GTG 
                 TTG 
                 V216L 
                   
                   
                   
                 Non 
                 7.00E−04 
                 Yes 
                 7 
                 13 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12715 
                 216 
                 GTG 
                 CTG 
                 V216L 
                   
                   
                   
                 Non 
                 7.00E−04 
                 Yes 
                 7 
                 1 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12716 
                 216 
                 GTG 
                 GAG 
                 V216E 
                   
                   
                   
                 Non 
                 7.00E−04 
                 Yes 
                 4 
                 7 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12716 
                 216 
                 GTG 
                 GGG 
                 V216G 
                   
                   
                   
                 Non 
                 7.00E−04 
                 Yes 
                 3 
                 5 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12727 
                 220 
                 TAT 
                 AAT 
                 Y220N 
                   
                   
                   
                 Non 
                 4.00E−06 
                 Yes 
                 12 
                 17 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12727 
                 220 
                 TAT 
                 CAT 
                 Y220H 
                   
                   
                   
                 Non 
                 4.00E−06 
                 Yes 
                 7 
                 16 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12727 
                 220 
                 TAT 
                 GAT 
                 Y220D 
                   
                   
                   
                 Non 
                 4.00E−06 
                 Yes 
                 2 
                 3 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12728 
                 220 
                 TAT 
                 TGT 
                 Y220C 
                   
                   
                   
                 Non 
                 4.00E−06 
                 Yes 
                 186 
                 360 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12728 
                 220 
                 TAT 
                 TCT 
                 Y220S 
                   
                   
                   
                 Non 
                 4.00E−06 
                 Yes 
                 9 
                 13 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 12729 
                 220 
                 TAT 
                 TAA 
                 Y220* 
                 Yes 
                   
                   
                   
                 4.00E−06 
                 Yes 
                   
                 2 
                 4 
               
               
                 12729 
                 220 
                 TAT 
                 TAG 
                 Y220* 
                 Yes 
                   
                   
                   
                 4.00E−06 
                 Yes 
                   
                 3 
                 4 
               
               
                 12730 
                 221 
                 GAG 
                 TAG 
                 E221* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 11 
                 3.5 
               
               
                 12739 
                 224 
                 GAG 
                 TAG 
                 E224* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 10 
                 3 
               
               
                 13324 
                 229 
                 TGT 
                 TGA 
                 C229* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 7 
                 3 
               
               
                 13337 
                 234 
                 TAC 
                 CAC 
                 Y234H 
                   
                   
                   
                 Non 
                 2.00E−01 
                   
                 12 
                 27 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13338 
                 234 
                 TAC 
                 TGC 
                 Y234C 
                   
                   
                   
                 Non 
                 2.00E−01 
                   
                 65 
                 136 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13339 
                 234 
                 TAC 
                 TAA 
                 Y234* 
                 Yes 
                   
                   
                   
                 2.00E−01 
                   
                   
                 11 
                 3.5 
               
               
                 13339 
                 234 
                 TAC 
                 TAG 
                 Y234* 
                 Yes 
                   
                   
                   
                 2.00E−01 
                   
                   
                 1 
                 3 
               
               
                 13343 
                 236 
                 TAC 
                 GAC 
                 Y236D 
                   
                   
                 Yes 
                 Non 
                 1.00E−02 
                 Yes 
                 5 
                 8 
                 4 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13343 
                 236 
                 TAC 
                 CAC 
                 Y236H 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 7 
                 14 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13343 
                 236 
                 TAC 
                 AAC 
                 Y236N 
                   
                   
                   
                   
                 1.00E−02 
                 Yes 
                 11 
                 19 
                 2 
               
               
                 13344 
                 236 
                 TAC 
                 TGC 
                 Y236C 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 41 
                 80 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13344 
                 236 
                 TAC 
                 TCC 
                 Y236S 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 3 
                 3 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13345 
                 236 
                 TAC 
                 TAA 
                 Y236* 
                 Yes 
                   
                   
                   
                 1.00E−02 
                 Yes 
                   
                 14 
                 4.5 
               
               
                 13345 
                 236 
                 TAC 
                 TAG 
                 Y236* 
                 Yes 
                   
                   
                   
                 1.00E−02 
                 Yes 
                   
                 8 
                 4 
               
               
                 13348 
                 237 
                 ATG 
                 ATA 
                 M237I 
                   
                   
                 Yes 
                 Non 
                 1.00E−01 
                   
                 1 
                 121 
                 3.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13348 
                 237 
                 ATG 
                 ATT 
                 M237I 
                   
                   
                 Yes 
                 Non 
                 1.00E−01 
                   
                 1 
                 50 
                 3.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13348 
                 237 
                 ATG 
                 ATC 
                 M237I 
                   
                   
                 Yes 
                 Non 
                 1.00E−01 
                   
                 1 
                 13 
                 3.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13349 
                 238 
                 TGT 
                 CGT 
                 C238R 
                   
                   
                   
                 Non 
                 6.00E−01 
                   
                 14 
                 23 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13350 
                 238 
                 TGT 
                 TAT 
                 C238Y 
                   
                   
                 Yes 
                 Non 
                 6.00E−01 
                   
                 42 
                 85 
                 4 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13350 
                 238 
                 TGT 
                 TTT 
                 C238F 
                   
                   
                 Yes 
                 Non 
                 6.00E−01 
                   
                 26 
                 40 
                 4 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13351 
                 238 
                 TGT 
                 TGA 
                 C238* 
                 Yes 
                   
                   
                   
                 6.00E−01 
                   
                   
                 7 
                 3 
               
               
                 13352 
                 239 
                 AAC 
                 GAC 
                 N239D 
                   
                   
                   
                 Non 
                   
                   
                 25 
                 48 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13353 
                 239 
                 AAC 
                 AGC 
                 N239S 
                   
                   
                 Yes 
                 Non 
                   
                   
                 1 
                 31 
                 3.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13355 
                 240 
                 AGT 
                 GGT 
                 S240G 
                   
                   
                   
                 Non 
                   
                   
                 13 
                 18 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13358 
                 241 
                 TCC 
                 ACC 
                 S241T 
                   
                   
                 Yes 
                 Non 
                 3.00E−06 
                 Yes 
                 5 
                 9 
                 4 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13358 
                 241 
                 TCC 
                 GCC 
                 S241A 
                   
                   
                   
                 Non 
                 3.00E−06 
                 Yes 
                 6 
                 12 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13358 
                 241 
                 TCC 
                 CCC 
                 S241P 
                   
                   
                   
                 Non 
                 3.00E−06 
                 Yes 
                 3 
                 10 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13359 
                 241 
                 TCC 
                 TTC 
                 S241F 
                   
                   
                 Yes 
                 Non 
                 3.00E−06 
                 Yes 
                 5 
                 100 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13359 
                 241 
                 TCC 
                 TGC 
                 S241C 
                   
                   
                   
                 Non 
                 3.00E−06 
                 Yes 
                 23 
                 35 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13359 
                 241 
                 TCC 
                 TAC 
                 S241Y 
                   
                   
                   
                 Non 
                 3.00E−06 
                 Yes 
                 6 
                 19 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13361 
                 242 
                 TGC 
                 AGC 
                 C242S 
                   
                   
                   
                 Non 
                 6.00E−03 
                 Yes 
                 10 
                 14 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13361 
                 242 
                 TGC 
                 CGC 
                 C242R 
                   
                   
                   
                 Non 
                 6.00E−03 
                 Yes 
                 10 
                 13 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13361 
                 242 
                 TGC 
                 GGC 
                 C242G 
                   
                   
                   
                 Non 
                 6.00E−03 
                 Yes 
                 2 
                 7 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13362 
                 242 
                 TGC 
                 TAC 
                 C242Y 
                   
                   
                 Yes 
                 Non 
                 6.00E−03 
                 Yes 
                 35 
                 54 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13362 
                 242 
                 TGC 
                 TTC 
                 C242F 
                   
                   
                 Yes 
                 Non 
                 6.00E−03 
                 Yes 
                 1 
                 88 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13362 
                 242 
                 TGC 
                 TCC 
                 C242S 
                   
                   
                   
                 Non 
                 6.00E−03 
                 Yes 
                 10 
                 19 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13363 
                 242 
                 TGC 
                 TGA 
                 C242* 
                 Yes 
                   
                   
                   
                 6.00E−03 
                 Yes 
                   
                 4 
                 4 
               
               
                 13363 
                 242 
                 TGC 
                 TGG 
                 C242W 
                   
                   
                   
                 Non 
                 6.00E−03 
                 Yes 
                 7 
                 13 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13367 
                 244 
                 GGC 
                 AGC 
                 G244S 
                   
                   
                 Yes 
                 Non 
                 4.00E−02 
                 Yes 
                 34 
                 70 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13367 
                 244 
                 GGC 
                 TGC 
                 G244C 
                   
                   
                   
                 Non 
                 4.00E−02 
                 Yes 
                 30 
                 47 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13367 
                 244 
                 GGC 
                 CGC 
                 G244R 
                   
                   
                   
                 Non 
                 4.00E−02 
                 Yes 
                 3 
                 5 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13368 
                 244 
                 GGC 
                 GAC 
                 G244D 
                   
                   
                 Yes 
                 Non 
                 4.00E−02 
                 Yes 
                 31 
                 63 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13368 
                 244 
                 GGC 
                 GTC 
                 G244V 
                   
                   
                   
                 Non 
                 4.00E−02 
                 Yes 
                 13 
                 24 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13368 
                 244 
                 GGC 
                 GCC 
                 G244A 
                   
                   
                   
                 Non 
                 4.00E−02 
                 Yes 
                 9 
                 11 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13370 
                 245 
                 GGC 
                 AGC 
                 G245S 
                   
                   
                 Yes 
                 Non 
                 6.00E−06 
                 Yes 
                 258 
                 436 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13370 
                 245 
                 GGC 
                 CGC 
                 G245R 
                   
                   
                 Yes 
                 Non 
                 6.00E−06 
                 Yes 
                 9 
                 18 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13370 
                 245 
                 GGC 
                 TGC 
                 G245C 
                   
                   
                   
                 Non 
                 6.00E−06 
                 Yes 
                 43 
                 83 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13371 
                 245 
                 GGC 
                 GAC 
                 G245D 
                   
                   
                 Yes 
                 Non 
                 6.00E−06 
                 Yes 
                 83 
                 153 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13371 
                 245 
                 GGC 
                 GTC 
                 G245V 
                   
                   
                   
                 Non 
                 6.00E−06 
                 Yes 
                 1 
                 73 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13371 
                 245 
                 GGC 
                 GCC 
                 G245A 
                   
                   
                   
                 Non 
                 6.00E−06 
                 Yes 
                 7 
                 13 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13373 
                 246 
                 ATG 
                 GTG 
                 M246V 
                   
                   
                 Yes 
                 Non 
                 5.00E−02 
                   
                 25 
                 55 
                 4 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13373 
                 246 
                 ATG 
                 TTG 
                 M246L 
                   
                   
                 Yes 
                 Non 
                 5.00E−02 
                   
                 2 
                 7 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13373 
                 246 
                 ATG 
                 CTG 
                 M246L 
                   
                   
                 Yes 
                 Non 
                 5.00E−02 
                   
                 2 
                 1 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13374 
                 246 
                 ATG 
                 AGG 
                 M246R 
                   
                   
                 Yes 
                 Non 
                 5.00E−02 
                   
                 10 
                 14 
                 3.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13377 
                 247 
                 AAC 
                 ATC 
                 N247I 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 1 
                 7 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13377 
                 247 
                 AAC 
                 ACC 
                 N247T 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 4 
                 4 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13379 
                 248 
                 CGG 
                 TGG 
                 R248W 
                   
                   
                 Yes 
                 Non 
                 7.00E−17 
                 Yes 
                 402 
                 707 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13379 
                 248 
                 CGG 
                 GGG 
                 R248G 
                   
                   
                   
                 Non 
                 7.00E−17 
                 Yes 
                 10 
                 23 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13380 
                 248 
                 CGG 
                 CAG 
                 R248Q 
                   
                   
                 Yes 
                 Non 
                 7.00E−17 
                 Yes 
                 5 
                 865 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13380 
                 248 
                 CGG 
                 CTG 
                 R248L 
                   
                   
                 Yes 
                 Non 
                 7.00E−17 
                 Yes 
                 2 
                 114 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13380 
                 248 
                 CGG 
                 CCG 
                 R248P 
                   
                   
                   
                 Non 
                 7.00E−17 
                 Yes 
                 11 
                 18 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13382 
                 249 
                 AGG 
                 TGG 
                 R249W 
                   
                   
                 Yes 
                 Non 
                 2.00E−03 
                 Yes 
                 23 
                 40 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13382 
                 249 
                 AGG 
                 GGG 
                 R249G 
                   
                   
                   
                 Non 
                 2.00E−03 
                 Yes 
                 23 
                 47 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13383 
                 249 
                 AGG 
                 ATG 
                 R249M 
                   
                   
                 Yes 
                 Non 
                 2.00E−03 
                 Yes 
                 25 
                 66 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13383 
                 249 
                 AGG 
                 AAG 
                 R249K 
                   
                   
                   
                 Non 
                 2.00E−03 
                 Yes 
                 14 
                 27 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13383 
                 249 
                 AGG 
                 ACG 
                 R249T 
                   
                   
                   
                 Non 
                 2.00E−03 
                 Yes 
                 15 
                 30 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13384 
                 249 
                 AGG 
                 AGT 
                 R249S 
                   
                   
                 Yes 
                 Non 
                 2.00E−03 
                 Yes 
                 282 
                 389 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13384 
                 249 
                 AGG 
                 AGC 
                 R249S 
                   
                   
                 Yes 
                 Non 
                 2.00E−03 
                 Yes 
                 282 
                 40 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13387 
                 250 
                 CCC 
                 CTC 
                 P250L 
                   
                   
                   
                 Non 
                 2.00E−03 
                 Yes 
                 51 
                 10 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13400 
                 255 
                 ATC 
                 TTC 
                 I255F 
                   
                   
                   
                 Non 
                   
                   
                 15 
                 39 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13407 
                 257 
                 CTG 
                 CAG 
                 L257Q 
                   
                   
                 Yes 
                 Non 
                   
                   
                 6 
                 16 
                 3.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13407 
                 257 
                 CTG 
                 CCG 
                 L257P 
                   
                   
                 Yes 
                 Non 
                   
                   
                 7 
                 14 
                 3.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13409 
                 258 
                 GAA 
                 TAA 
                 E258* 
                 Yes 
                   
                   
                   
                 8.00E−02 
                 Yes 
                   
                 25 
                 3.5 
               
               
                 13409 
                 258 
                 GAA 
                 AAA 
                 E258K 
                   
                   
                   
                 Non 
                 2.00E−03 
                 Yes 
                 67 
                 14 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13412 
                 259 
                 GAC 
                 TAC 
                 D259Y 
                   
                   
                 Yes 
                 Non 
                 7.00E−02 
                 Yes 
                 14 
                 32 
                 4 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13413 
                 259 
                 GAC 
                 GTC 
                 D259V 
                   
                   
                   
                 Non 
                 7.00E−02 
                 Yes 
                 12 
                 19 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13774 
                 265 
                 CTG 
                 CCG 
                 L265P 
                   
                   
                 Yes 
                 Non 
                   
                   
                 1 
                 20 
                 3.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13776 
                 266 
                 GGA 
                 TGA 
                 G266* 
                 Yes 
                   
                   
                   
                 2.00E−02 
                 Yes 
                   
                 29 
                 4.5 
               
               
                 13776 
                 266 
                 GGA 
                 AGA 
                 G266R 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 14 
                 51 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13776 
                 266 
                 GGA 
                 CGA 
                 G266R 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 14 
                 18 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13777 
                 266 
                 GGA 
                 GAA 
                 G266E 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 41 
                 79 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13777 
                 266 
                 GGA 
                 GTA 
                 G266V 
                   
                   
                   
                 Non 
                 2.00E−02 
                 Yes 
                 22 
                 55 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13779 
                 267 
                 CGG 
                 TGG 
                 R267W 
                   
                   
                   
                 Non 
                   
                   
                 19 
                 31 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13780 
                 267 
                 CGG 
                 CCG 
                 R267P 
                   
                   
                   
                 Non 
                   
                   
                 12 
                 17 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13789 
                 270 
                 TTT 
                 TGT 
                 F270C 
                   
                   
                   
                 Non 
                 3.00E−01 
                   
                 15 
                 27 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13791 
                 271 
                 GAG 
                 TAG 
                 E271* 
                 Yes 
                   
                   
                   
                 5.00E−02 
                   
                   
                 17 
                 3.5 
               
               
                 13794 
                 272 
                 GTG 
                 ATG 
                 V272M 
                   
                   
                   
                 Non 
                 1.00E−01 
                   
                 60 
                 103 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13797 
                 273 
                 CGT 
                 TGT 
                 R273C 
                   
                   
                 Yes 
                 Non 
                 4.00E−18 
                 Yes 
                 363 
                 664 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13797 
                 273 
                 CGT 
                 AGT 
                 R273S 
                   
                   
                   
                 Non 
                 4.00E−18 
                 Yes 
                 1 
                 19 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13797 
                 273 
                 CGT 
                 GGT 
                 R273G 
                   
                   
                   
                 Non 
                 4.00E−18 
                 Yes 
                 8 
                 17 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13798 
                 273 
                 CGT 
                 CAT 
                 R273H 
                   
                   
                 Yes 
                 Non 
                 4.00E−18 
                 Yes 
                 434 
                 812 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13798 
                 273 
                 CGT 
                 CTT 
                 R273L 
                   
                   
                 Yes 
                 Non 
                 4.00E−18 
                 Yes 
                 79 
                 142 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13798 
                 273 
                 CGT 
                 CCT 
                 R273P 
                   
                   
                   
                 Non 
                 4.00E−18 
                 Yes 
                 22 
                 37 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13800 
                 274 
                 GTT 
                 TTT 
                 V274F 
                   
                   
                 Yes 
                 Non 
                 7.00E−01 
                   
                 14 
                 33 
                 4 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13801 
                 274 
                 GTT 
                 GCT 
                 V274A 
                   
                   
                   
                 Non 
                 7.00E−01 
                   
                 11 
                 18 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13803 
                 275 
                 TGT 
                 CGT 
                 C275R 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 6 
                 15 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13803 
                 275 
                 TGT 
                 AGT 
                 C275S 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 2 
                 0 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13803 
                 275 
                 TGT 
                 GGT 
                 C275G 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 7 
                 8 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13804 
                 275 
                 TGT 
                 TAT 
                 C275Y 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 40 
                 75 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13804 
                 275 
                 TGT 
                 TTT 
                 C275F 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 30 
                 49 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13804 
                 275 
                 TGT 
                 TCT 
                 C275S 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 2 
                 2 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13805 
                 275 
                 TGT 
                 TGA 
                 C275* 
                 Yes 
                   
                   
                   
                 1.00E−02 
                 Yes 
                   
                 3 
                 4 
               
               
                 13805 
                 275 
                 TGT 
                 TGG 
                 C275W 
                   
                   
                   
                 Non 
                 1.00E−02 
                 Yes 
                 7 
                 10 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13806 
                 276 
                 GCC 
                 CCC 
                 A276P 
                   
                   
                 Yes 
                 Non 
                 7.00E−02 
                   
                 11 
                 18 
                 4 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13810 
                 277 
                 TGT 
                 TAT 
                 C277Y 
                   
                   
                   
                 Non 
                 1.00E−01 
                   
                 14 
                 27 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13810 
                 277 
                 TGT 
                 TTT 
                 C277F 
                   
                   
                   
                 Non 
                 1.00E−01 
                   
                 19 
                 51 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13811 
                 277 
                 TGT 
                 TGA 
                 C277* 
                 Yes 
                   
                   
                   
                 1.00E−01 
                   
                   
                 8 
                 3 
               
               
                 13812 
                 278 
                 CCT 
                 ACT 
                 P278T 
                   
                   
                   
                 Non 
                 1.00E−05 
                 Yes 
                 21 
                 32 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13812 
                 278 
                 CCT 
                 TCT 
                 P278S 
                   
                   
                   
                 Non 
                 1.00E−05 
                 Yes 
                 46 
                 82 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13812 
                 278 
                 CCT 
                 GCT 
                 P278A 
                   
                   
                   
                 Non 
                 1.00E−05 
                 Yes 
                 16 
                 28 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13813 
                 278 
                 CCT 
                 CAT 
                 P278H 
                   
                   
                 Yes 
                 Non 
                 1.00E−05 
                 Yes 
                 10 
                 14 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13813 
                 278 
                 CCT 
                 CTT 
                 P278L 
                   
                   
                   
                 Non 
                 1.00E−05 
                 Yes 
                 51 
                 81 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13813 
                 278 
                 CCT 
                 CGT 
                 P278R 
                   
                   
                   
                 Non 
                 1.00E−05 
                 Yes 
                 24 
                 42 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13816 
                 279 
                 GGG 
                 GAG 
                 G279E 
                   
                   
                 Yes 
                 Non 
                   
                   
                 1 
                 45 
                 3.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13818 
                 280 
                 AGA 
                 TGA 
                 R280* 
                 Yes 
                   
                   
                   
                 2.00E−01 
                   
                   
                 9 
                 3 
               
               
                 13818 
                 280 
                 AGA 
                 GGA 
                 R280G 
                   
                   
                   
                 Non 
                 2.00E−01 
                   
                 15 
                 41 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13819 
                 280 
                 AGA 
                 ACA 
                 R280T 
                   
                   
                 Yes 
                 Non 
                 2.00E−01 
                   
                 46 
                 91 
                 4 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13819 
                 280 
                 AGA 
                 AAA 
                 R280K 
                   
                   
                   
                 Non 
                 2.00E−01 
                   
                 37 
                 70 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13819 
                 280 
                 AGA 
                 ATA 
                 R280I 
                   
                   
                   
                 Non 
                 2.00E−01 
                   
                 11 
                 24 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13820 
                 280 
                 AGA 
                 AGT 
                 R280S 
                   
                   
                 Yes 
                 Non 
                 2.00E−01 
                   
                 4 
                 19 
                 3.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13820 
                 280 
                 AGA 
                 AGC 
                 R280S 
                   
                   
                 Yes 
                 Non 
                 2.00E−01 
                   
                 4 
                 5 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13821 
                 281 
                 GAC 
                 AAC 
                 D281N 
                   
                   
                 Yes 
                 Non 
                 4.00E−03 
                 Yes 
                 18 
                 33 
                 5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13821 
                 281 
                 GAC 
                 TAC 
                 D281Y 
                   
                   
                 Yes 
                 Non 
                 4.00E−03 
                 Yes 
                 5 
                 11 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13821 
                 281 
                 GAC 
                 CAC 
                 D281H 
                   
                   
                   
                 Non 
                 4.00E−03 
                 Yes 
                 18 
                 38 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13822 
                 281 
                 GAC 
                 GGC 
                 D281G 
                   
                   
                 Yes 
                 Non 
                 4.00E−03 
                 Yes 
                 9 
                 14 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13822 
                 281 
                 GAC 
                 GTC 
                 D281V 
                   
                   
                   
                 Non 
                 4.00E−03 
                 Yes 
                 2 
                 4 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13822 
                 281 
                 GAC 
                 GCC 
                 D281A 
                   
                   
                   
                 Non 
                 4.00E−03 
                 Yes 
                 2 
                 5 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13823 
                 281 
                 GAC 
                 GAA 
                 D281E 
                   
                   
                 Yes 
                 Non 
                 4.00E−03 
                 Yes 
                 9 
                 20 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13823 
                 281 
                 GAC 
                 GAG 
                 D281E 
                   
                   
                 Yes 
                 Non 
                 4.00E−03 
                 Yes 
                 9 
                 24 
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13824 
                 282 
                 CGG 
                 GGG 
                 R282G 
                   
                   
                   
                 Non 
                 2.00E−07 
                 Yes 
                 25 
                 46 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13824 
                 282 
                 CGG 
                 TGG 
                 R282W 
                   
                   
                   
                 Non 
                 2.00E−07 
                 Yes 
                 10 
                 554 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13825 
                 282 
                 CGG 
                 CCG 
                 R282P 
                   
                   
                   
                 Non 
                 2.00E−07 
                 Yes 
                 13 
                 22 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13825 
                 282 
                 CGG 
                 CAG 
                 R282Q 
                   
                   
                   
                   
                 2.00E−07 
                 Yes 
                 19 
                 26 
                 2 
               
               
                 13828 
                 283 
                 CGC 
                 CAC 
                 R283H 
                   
                   
                   
                 Non 
                 5.00E−01 
                   
                 11 
                 17 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13828 
                 283 
                 CGC 
                 CCC 
                 R283P 
                   
                   
                   
                 Non 
                 5.00E−01 
                   
                 21 
                 35 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13833 
                 285 
                 GAG 
                 TAG 
                 E285* 
                 Yes 
                   
                   
                   
                 1.00E−01 
                   
                   
                 21 
                 3.5 
               
               
                 13833 
                 285 
                 GAG 
                 AAG 
                 E285K 
                   
                   
                   
                 Non 
                 1.00E−01 
                   
                 87 
                 169 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13834 
                 285 
                 GAG 
                 GTG 
                 E285V 
                   
                   
                   
                 Non 
                 1.00E−01 
                   
                 13 
                 19 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13836 
                 286 
                 GAA 
                 TAA 
                 E286* 
                 Yes 
                   
                   
                   
                 1.00E−03 
                 Yes 
                   
                 21 
                 4.5 
               
               
                 13836 
                 286 
                 GAA 
                 AAA 
                 E286K 
                   
                   
                   
                 Non 
                 1.00E−03 
                 Yes 
                 1 
                 83 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13836 
                 286 
                 GAA 
                 CAA 
                 E286Q 
                   
                   
                   
                 Non 
                 1.00E−03 
                 Yes 
                 5 
                 15 
                 2.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13837 
                 286 
                 GAA 
                 GGA 
                 E286G 
                   
                   
                   
                 Non 
                 1.00E−03 
                 Yes 
                 14 
                 19 
                 3 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13837 
                 286 
                 GAA 
                 GTA 
                 E286V 
                   
                   
                   
                 Non 
                 1.00E−03 
                 Yes 
                 6 
                 6 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13837 
                 286 
                 GAA 
                 GCA 
                 E286A 
                   
                   
                   
                 Non 
                 1.00E−03 
                 Yes 
                 1 
                 2 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13838 
                 286 
                 GAA 
                 GAT 
                 E286D 
                   
                   
                   
                 Non 
                 1.00E−03 
                 Yes 
                 1 
                 3 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13838 
                 286 
                 GAA 
                 GAC 
                 E286D 
                   
                   
                   
                 Non 
                 1.00E−03 
                 Yes 
                 1 
                 1 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 13839 
                 287 
                 GAG 
                 TAG 
                 E287* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 14 
                 3.5 
               
               
                 13842 
                 288 
                 AAT 
                 GAT 
                 N288D 
                   
                   
                 Yes 
                   
                   
                   
                 1 
                 1 
                 2 
               
               
                 13851 
                 291 
                 AAG 
                 TAG 
                 K291* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 7 
                 3 
               
               
                 13854 
                 292 
                 AAA 
                 TAA 
                 K292* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 2 
                 3 
               
               
                 13860 
                 294 
                 GAG 
                 TAG 
                 E294* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 57 
                 3.5 
               
               
                 13872 
                 298 
                 GAG 
                 TAG 
                 E298* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 64 
                 3.5 
               
               
                 13893 
                 305 
                 AAG 
                 TAG 
                 K305* 
                 Yes 
                   
                   
                   
                   
                   
                   
                 15 
                 3.5 
               
               
                 13896 
                 306 
                 CGA 
                 TGA 
                 R306* 
                 Yes 
                   
                   
                 Non 
                 9.00E−02 
                   
                 164 
                   
                 4.5 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                 16900 
                 337 
                 CGC 
                 TGC 
                 R337C 
                   
                   
                   
                 Non 
                 9.00E−02 
                   
                 11 
                 18 
                 2 
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                 functional 
               
               
                   
               
             
          
           
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 P53 
                 Reported &gt;10x 
                 Reported &gt;10x 
                   
               
               
                   
                   
                   
                   
                   
                   
                   
                   
                   
                 signature 
                 in somatic 
                 in somatic 
               
               
                   
                   
                   
                   
                 Stop 
                 Splice 
                 Dominant 
                 Trans- 
                 P53 
                 p value &lt; 
                 @ AA 
                 @ Base 
                 Evi- 
               
               
                   
                   
                   
                 Protein 
                 Codon 
                 Defect 
                 Negative 
                 activation 
                 Signature 
                 0.05 
                 level 
                 level 
                 dence 
               
               
                 NtNum 
                 Codon 
                 Change 
                 Change 
                 (3 pts) 
                 (3 pts) 
                 (2 pts) 
                 (1 pt) 
                 P-Value 
                 (1 pt) 
                 (0.5 pt) 
                 (0.5 pt) 
                 Score 
               
               
                   
               
               
                 11080 
                 2-intron 
                 G &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11080 
                 2-intron 
                 G &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11080 
                 2-intron 
                 G &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11081 
                 2-intron 
                 del167 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11081 
                 2-intron 
                 T &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11081 
                 2-intron 
                 T &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11081 
                 2-intron 
                 T &gt; G 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11195 
                 2-intron 
                 A &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11195 
                 2-intron 
                 A &gt; G 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11195 
                 2-intron 
                 A &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11196 
                 2-intron 
                 G &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11196 
                 2-intron 
                 G &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11196 
                 2-intron 
                 G &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11219 
                 3-intron 
                 G &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11219 
                 3-intron 
                 G &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11219 
                 3-intron 
                 G &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11220 
                 3-intron 
                 T &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11220 
                 3-intron 
                 T &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11220 
                 3-intron 
                 T &gt; G 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11326 
                 3-intron 
                 A &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11326 
                 3-intron 
                 A &gt; G 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11326 
                 3-intron 
                 A &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11327 
                 3-intron 
                 del19 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11327 
                 3-intron 
                 G &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11327 
                 3-intron 
                 G &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11327 
                 3-intron 
                 G &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11607 
                 4-intron 
                 G &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11607 
                 4-intron 
                 G &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11607 
                 4-intron 
                 G &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11608 
                 4-intron 
                 ins1 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11608 
                 4-intron 
                 T &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11608 
                 4-intron 
                 T &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 11608 
                 4-intron 
                 T &gt; G 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12362 
                 4-intron 
                 A &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12362 
                 4-intron 
                 A &gt; G 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12362 
                 4-intron 
                 A &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12362 
                 4-intron 
                 del1 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12362 
                 4-intron 
                 ins1 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12363 
                 4-intron 
                 del1 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12363 
                 4-intron 
                 G &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12363 
                 4-intron 
                 G &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12363 
                 4-intron 
                 G &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12548 
                 5-intron 
                 G &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12548 
                 5-intron 
                 G &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12548 
                 5-intron 
                 G &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12549 
                 5-intron 
                 T &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12549 
                 5-intron 
                 T &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12549 
                 5-intron 
                 T &gt; G 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12627 
                 5-intron 
                 A &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12627 
                 5-intron 
                 A &gt; G 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12627 
                 5-intron 
                 A &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12627 
                 5-intron 
                 del10 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12628 
                 5-intron 
                 del1 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12628 
                 5-intron 
                 G &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12628 
                 5-intron 
                 G &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12628 
                 5-intron 
                 G &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12628 
                 5-intron 
                 GG &gt; AA 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12742 
                 6-intron 
                 del1 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12742 
                 6-intron 
                 G &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12742 
                 6-intron 
                 G &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12742 
                 6-intron 
                 G &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12743 
                 6-intron 
                 T &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12743 
                 6-intron 
                 T &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 12743 
                 6-intron 
                 T &gt; G 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13308 
                 6-intron 
                 A &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13308 
                 6-intron 
                 A &gt; G 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13308 
                 6-intron 
                 A &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13308 
                 6-intron 
                 del11 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13309 
                 6-intron 
                 G &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13309 
                 6-intron 
                 G &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13309 
                 6-intron 
                 G &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13420 
                 7-intron 
                 del137 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13420 
                 7-intron 
                 del342(del 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                   
                   
                 intron7) 
               
               
                 13420 
                 7-intron 
                 G &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13420 
                 7-intron 
                 G &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13420 
                 7-intron 
                 G &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13421 
                 7-intron 
                 del10 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13421 
                 7-intron 
                 T &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13421 
                 7-intron 
                 T &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13421 
                 7-intron 
                 T &gt; G 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13759 
                 7-intron 
                 G &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13761 
                 7-intron 
                 A &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13761 
                 7-intron 
                 A &gt; G 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13761 
                 7-intron 
                 A &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13762 
                 7-intron 
                 G &gt; A 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13762 
                 7-intron 
                 G &gt; C 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13762 
                 7-intron 
                 G &gt; T 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                 13762 
                 7-intron 
                 ins3 
                 N/A 
                   
                 Yes 
                   
                   
                   
                   
                   
                   
                 3 
               
               
                   
               
               
                 “del” denotes deletion of the number of basepairs; “ins” denotes insertion of the number of baspairs. 
               
             
          
         
       
     
       Example 2 
       [0128]    A Phase II study was conducted with WEE1-1 Inhibitor combined with carboplatin in patients with p53 mutated epithelial ovarian cancer that show early relapse (&lt;3 months) or progression during standard first line treatment with carboplatin—paclitaxel combination therapy. Patients were enrolled based on the tumor&#39;s TP53 gene sequence as determined in the AmpliChip p53 assay. 
         [0129]    AmpliChip p53 test reagents are used to amplify products encompassing the coding regions of the p53 gene in two reactions (A and B) for all samples including a reference wildtype DNA. Exons 2, 5, 8, and 10; exon 4 upstream sequence; and internal control are in the Primer Mix A. Primer Mix B is designed and contained primers for exons 3, 6, 7, 9, and 11; exon 4 downstream sequence; and internal control. After thermal cycling, the products from Primer Mixes A and B are combined. The products generated from the A and B reactions are cleaved by a mix containing DNase I. Fragmentation is performed by recombinant DNase I to generate small DNA fragments of an average size of 50-100 nucleotides. The alkaline phosphatase in the Working Fragmentation Mix destroys the residual dNTPs from the amplification reactions. The fragmented DNA amplicons are subsequently labeled with biotin at their 3′ termini by the action of terminal transferase, using AmpliChip TdT Labeling Reagent as substrate. The biotin-labeled p53 target DNA fragments are added to the hybridization buffer containing the AmpliChip Oligonucleotide Solution which functions as a hybridization control. The mixture is hybridized to the oligonucleotides located on the AmpliChip p53 Microarray using the Affymetrix GeneChip Fluidics Station 450Dx and an AmpliChip p53 specific protocol. The hybridized AmpliChip p53 Microarray is washed and stained with a streptavidin-conjugated fluorescent dye. 
       Design of the AmpliChip p53 Microarray 
       [0130]    The microarray consists of a square grid of 228,484 probes, with sides that are 11 micron each. Each probe contains multiple copies of a specific oligonucleotide sequence. A single probe set for an interrogating base position includes five probes, one probe to hybridize to the wild type, three probes to detect three possible single base pair mutations, and one probe to detect single deletion. There are at least 24 probe sets for each nucleotide position, including both sense and antisense probe sequences. A total of 1300 nucleotide positions of coding regions of exons 2-11 are tiled on AmpliChip p53. AmpliChip p53 Microarrays are manufactured using technology that combines photolithographic methods and combinatorial chemistry. Over 220,000 different oligonucleotide probes are synthesized on a glass surface to analyze both sense and antisense strands of an amplified target DNA specimen. Within the 11×11 μm 2  probe microarray, each probe type is located in a specific area called a probe cell, which contains approximately 106 copies of a given probe. Probe microarrays are manufactured by light-directed combinatorial chemistry in a series of cycles. The glass substrates are coated with linkers containing photolabile protecting groups. A mask is then applied that exposes selected portions of the probe microarray. Illumination removes the photolabile protecting groups enabling selective nucleoside phosphoramidite addition only at the previously exposed sites. Next, a different mask is applied and the cycle of illumination and chemical coupling is performed again. By repeating this cycle, a specific set of oligonucleotide probes is synthesized, with each probe type in a known location. The completed probe microarrays are packaged into cartridges compatible with the GeneChip Fluidics Station 450Dx. After staining, the AmpliChip p53 Microarray is scanned by an Affymetrix GeneChip Scanner 3000Dx using a laser that excites the fluorescent label bound to the hybridized p53 target DNA fragments. The amount of emitted light is proportional to bound target DNA at each location on the probe microarray. 
       Data Analysis of Microarray Signals. 
       [0131]    The p53 mutation status is determined by a p53 mutation detection algorithm, which is designed to detect single base pair substitutions and single base pair deletions of a sample in a background of wild type p53 DNA probe intensities. The algorithm first reads the probe intensities generated by the GeneChip Operating Software, Version 1.1 provided by Affymetrix. Based on the raw data, the algorithm performs an initial exon quality test to detect distinct problems in each PCR product. If an exon fails the initial quality test, the exon failure is reported and no further analysis is made. If an exon passes the test, the probe intensities are normalized by using quantile normalization in order to correct array-to-array variability. 
         [0132]    The quality of each probe set is then examined to eliminate unreliable probe set data for further computation. Using probes sets that passed the quality tests, the algorithm makes a tentative call for each base position. Possible base calls for each nucleotide position are wild type, single base substitution, single base deletion, or no call (unable to make a call). In the case of a single base substitution, the algorithm identifies the mutated base (e.g., G®A). After the tentative calls are made, the reliability of each base call is reexamined by the algorithm to fine tune the calls using various parameters calculated from the neighboring base positions. Each exon quality is also reexamined based on the final base calls. If there are too many no calls and/or mutation calls in one exon, the data is considered as “noisy,” and the exon fails the quality test. If an exon fails, the exon failure is reported, and no calls are reported for that exon. 
       Tumor Response Evaluation 
       [0133]    Patients were evaluable for response to study treatment if at least one follow-up examination was performed at the end of the second treatment cycle (at least a six-week period). Tumor response was assessed either in measurable or evaluable tumor lesions according to the RECIST 1.0 criteria (Appendix IV). In the case of stable disease, a confirmation was necessary within 6 to 8 weeks of initial assessment. In case of stable disease at the end of treatment or in case of discontinuation for unacceptable adverse experiences evaluation took place every 2 months and CA-125 (cancer antigen) was determined. In case of a CA-125 increase a CT scan was performed. In patients for whom CA-125 is not a good marker, a CT-scan was performed every 2 months, until disease progression. Patients without radiologically measurable disease had elevated CA-125 prior to start and were evaluated based on CA-125 levels. CT-scans were also performed in these patients. According to the Gynecologic Cancer Intergroup (GCIG) CA-125 response criteria, progressive disease after a complete response to primary therapy is defined as follows: the date of first elevation of CA-125 to two-fold the upper limit of normal (ULN) (documented on two occasions at least a week apart). For those with persistently elevated CA-125 levels, progression of disease is defined as the first date of CA-125≧2× the nadir value documented on two occasions no less than a week apart. The ULN for CA-125 at the NKI is 35 U/mL. However, in this study for patients with CA-125 nadir ≦10 U/mL, a confirmed value of ≧20 U/mL served as an early signal of CA-125 progression, and for patients with nadir more than 10 U/mL, a value ≧2× nadir. The definition of disease progression included both the standard RECIST criterion and the CA-125 criterion as defined above, whichever occurs first. Response regarding CA-125 is defined standards as 50% reduction during treatment of CA-125. 
         [0134]    The following table summarizes the p53 amino acid mutation information for responders in the above clinical trial. 
         [0000]    
       
         
               
               
               
               
               
             
           
               
                 TABLE 4 
               
               
                   
               
               
                   
                   
                 Amino 
                   
                   
               
               
                   
                   
                 Acid/Gene 
                 Mutation listed 
               
               
                 Patient Number 
                 PCR 
                 mutation 
                 in Table 3 
                 Responder 
               
               
                   
               
             
             
               
                 1003 
                 Exon 7 
                 C238F 
                 Yes 
                 Yes 
               
               
                 1008 
                 Exon 7 
                 R248W 
                 Yes 
                 Yes 
               
               
                 1010 
                 Exon 8 
                 E298 stop codon 
                 Yes 
                 Yes 
               
               
                 1011 
                 Exon 8 
                 R273L 
                 Yes 
                 Yes 
               
               
                 1015 
                 Exon 5 
                 V157 frameshift 
                 Yes 
                 Yes 
               
               
                   
               
             
          
         
       
     
       Example 4 
       [0135]    In a randomized, Phase II study evaluating WEE1-1 in combination with paclitaxel and carboplatin versus paclitaxel and carboplatin alone in adult patients with platinum sensitive p53 mutant ovarian cancer, patient enrollment was based on the presence of one or more mutations in Table 3. If the specimen has no mutations in the TP53 gene, or if it contains a mutation not listed on the Mutation Lookup Table 3, the patient will not be eligible for the study. If the specimen had at least one mutation that is listed in the Mutation Lookup Table 3, the patient was eligible for enrollment in the study. 
         [0136]    Analysis of patient tumor tissue samples using the AmpliChip p53 assay was performed by Caris Life Sciences (Phoenix, Ariz.). The AmpliChip p53 assay characterized each specimen as ‘Mutation Not Detected,’ Mutation Detected′ or ‘Test Invalid’. A base change to a synonymous codon is treated as a Mutation Not Detected call, since it does not alter the amino acid. There are seven single-nucleotide polymorphisms reported in the IARC database R15 (2010 release) within the tiled nucleotide positions of AmpliChip 53. While three do not result in amino acid changes (codons 34, 36 and 213), four others result in the following amino acid changes: P47S, P72R, V217M and G360A. These are all treated as Mutation Not Detected, and are not reported. If at least one mutation is detected, the sample is called “Mutation Detected” and the nucleotide change detected is reported, along with the corresponding amino acid change in the p53 protein. If no base changes are detected and the test is valid, the sample is called “Mutation Not Detected.” 
       F. 
       [0000]