CN103038638A

CN103038638A - Methods for predicting sensitivity to treatment with a targeted tyrosine kinase inhibitor

Info

Publication number: CN103038638A
Application number: CN201180033733XA
Authority: CN
Inventors: E.M.麦基根; P.安塞尔; B.L.多维尔; K.张; V.德瓦纳拉彦
Original assignee: AbbVie Inc
Current assignee: AbbVie Inc
Priority date: 2010-05-07
Filing date: 2011-05-04
Publication date: 2013-04-10
Also published as: JP2013525814A; EP2567232A1; MX2012012986A; CA2798441A1; WO2011140234A1

Abstract

Methods and kits for predicting the sensitivity of a cancer to treatment with a targeted tyrosine kinase inhibitor are disclosed.

Description

Be used for predicting the method to the susceptibility of using the target treatment with tyrosine kinase inhibitors

Relevant application information

The application requires the right of priority of the u.s. patent application serial number 61/332,545 of application on May 7th, 2010, and is attached to herein by causing with its integral body.

Technical field

This instructions relates generally to be estimated and/or treatment trouble or the doubtful experimenter who suffers from the tumprigenicity patient's condition, relate in particular to biomarker for the identification of the patient's who accepts particular medication purposes, and it allows monitoring to the patient of such therapeutic response.

Background of invention

The genetic heterogeneity of cancer is to make the complicated factor of the effective cancer drug of exploitation.The cancer that the traditional histopathologic classification of foundation is considered to single disease entity often discloses several genes group hypotype when standing the molecule profile analysis.In some cases, as if different genome hypotypes has functional dependency with the effect of some drugs.For example, the existence of the effect of some target on cancer medicine and genome signature (for example gene magnification) relevant (referring to, for example, T.J. Lynch etc., " Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib (being that non-small cell lung cancer is to the activated mutant in the EGF-R ELISA on reactive basis of Gefitinib) ", N. Engl. J. Med., 350:2129-2139,2004.).

Specific blood plasma and the blood serum designated object that can be used for patients with lung cancer is carried out subclassification also identified in clinical research.American National clinical biochemical association (National Association of Clinical Biochemistry) announced tumor markers the practice guideline of clinical middle use and suggestion (referring to The NACB Practice Guidelines and Recommendations for Use of Tumor Markers in the Clinic (tumor markers is in the NACB of clinical middle use practice guideline and suggestion). Lung Cancer Section 3P).The pattern that tumor markers discharges is relevant with the histology background of tumour and can disclose the histology component of mixing.Table 1 has been summarized the correlativity of mark CYFRA 21-1, CEA, NSE and ProGRP and tumor histology.

Table 1:

Histology	Before the treatment	Follow up a case by regular visits to after the treatment
			Unknown	CYFRA 21-1、CEA、NSE、ProGRP	After the operation: according to the histology in the terminal illness: use the preceding mark
Gland cancer	CYFRA 21-1 and CEA	CYFRA 21-1 and/or CEA
			Squamous cell carcinoma	CYFRA 21-1 and CEA (and SCCA)	CYFRA 21-1 and/or CEA (and/or SCCA)
Large cell carcinoma	CYFRA 21-1 and CEA	CYFRA 21-1 and/or CEA
			Small cell carcinoma	NSE and ProGRP	NSE and/or ProGRP

CEA, carcinomebryonic antigen; CYFRA 21-1, cytokeratin 19 fragment; NSE, neuronspecific enolase; ProGRP, the progastrin release peptide; SCCA, squamous cell carcinoma antigen

Although the special sign thing can be helpful to distinguishing different histological subtypes from the correlativity of the lung cancer of specific subclass, the functional importance of such mark generally is not understood well.

ABT-869 (Linifanib) [N-(4-(3-amino-lH-indazole-4-yl) phenyl)-N'-(2-fluoro-5-aminomethyl phenyl) urea]) be a kind of multiple receptor targeted tyrosine kinase inhibitor, it has shown all members (for example, the KDR IC that suppresses VEGF and pdgf receptor family ₅₀Value is 4 nM), and have less activity (IC for incoherent receptor tyrosine kinase, solubility tyrosine kinase and serine/threonine kinase ₅₀Value〉1 μ M).In addition, it shows strong antiproliferative and cells apoptosis to depending on mutant (composing type activation) FLT3 and the kinase whose tumour cell of KIT.Although its strong antitumor activity, many malignant cell types are with the ABT-869 refractory.The reason of resistance is unknown.

Because ABT-869 for the potential therapeutical uses of kinds cancer, needs to identify the diagnostic assay of following to those best patients of the acceptance of ABT-869 treatment.In addition, the needs that have the diagnostic method of the effect can be used for monitoring the ABT-869 treatment.The further needs of following mensuration that have the service marking thing, this mark can for example be measured in blood or the blood plasma part at the tissue sample that obtains easily.

Summary of the invention

The level of having found serum soluble fragment (CYFRA 21-1), cancer antigen 125 (CA125) and the carcinomebryonic antigen (CEA) of mark neuronspecific enolase (NSE), Cyfra21-1 is that experimenter's cancer is to the indication of the susceptibility that gives medicine ABT-869.Method as herein described and kit part is based on such discovery: following any or multiple indication is not with respect to having the relatively experimenter of level to any mark, and experimenter's cancer increases the susceptibility that gives ABT-869: the level that the level that the level that the level of NSE is lower than the predeterminated level of NSE, CA125 is lower than predeterminated level, the CEA of CA125 is higher than the predeterminated level of CEA, CYFRA 21-1 is lower than predeterminated level or its any combination of CYFRA 21-1.

Therefore, in one aspect, this instructions provides a kind of cancer subtend experimenter for the prediction experimenter to give the method for the susceptibility of ABT-869, the method may further comprise the steps: measure at least one and be selected from the level of following mark in available from experimenter's sample: neuronspecific enolase (NSE), the serum soluble fragment of Cyfra21-1 (CYFRA 21-1), cancer antigen 125 (CA125) and carcinomebryonic antigen (CEA), wherein following any indication is with respect to NSE, the level of CYFRA 21-1 or CA125 be higher than each mark predeterminated level the experimenter or be lower than the experimenter of the predeterminated level of each mark with respect to the level of CEA, experimenter's cancer increases the susceptibility that gives ABT-869: the level of NSE is lower than the predeterminated level of NSE, the level of CA125 is lower than the predeterminated level of CA125, the level of CEA is higher than the predeterminated level of CEA, the level of CYFRA 21-1 is lower than predeterminated level or its any combination of CYFRA 21-1.Cancer can be non-small cell lung cancer.The method can comprise, for example measures at least two level that is selected from following mark: NSE, CA125, CYFRA21-1 and CEA.The method can comprise the level of measuring NSE, CA125, CYFRA21-1 and CEA.The method can comprise in addition, for example produce experimenter's mark signature (signature) from the level of two or more marks, wherein have the mark signature indication of preassigned pattern with respect to the mark signature that does not have preassigned pattern, the experimenter increases the susceptibility that gives ABT-869.The method can comprise in addition by application class tree analyzes level comparison with the identical mark in the level of two or more marks in the sample and the control sample.Classification tree analysis can be finished by computer procedures.

On the other hand, the cancer that this instructions provides a kind of experimenter of prediction is to the method for the susceptibility that gives ABT-869, the method may further comprise the steps: measure the marker levels in the mark group that comprises NSE, CA125, CYFRA 21-1 and CEA in available from experimenter's sample, and with the predeterminated level of each marker levels in the sample and each mark relatively, wherein, each mark in the sample gives the susceptibility of ABT-869 with respect to the level indication cancer subtend experimenter of the predeterminated level of each mark.In the method, the level of each mark in the sample and the predeterminated level of each mark are relatively comprised: with each marker levels in marker levels and the reference sample relatively, wherein this reference sample comprises each mark of the level corresponding with the predeterminated level of each mark.Cancer can be non-small cell lung cancer.In the method, NSE level in experimenter's the sample can be, for example be lower than the predeterminated level of NSE, CA125 level in experimenter's the sample can be lower than the predeterminated level of CA125, CYFRA 21-1 level in experimenter's the sample can be lower than the predeterminated level of CYFRA 21-1, or the CEA level in experimenter's the sample is higher than the predeterminated level of CEA, perhaps can have any combination of all four kinds of situations.The method can comprise the mark signature that produces the experimenter from the level of one or more marks in addition, wherein have the mark signature indication of preassigned pattern with respect to the experimenter with the mark signature that does not have preassigned pattern, the experimenter increases the susceptibility that gives ABT-869.The method can comprise in addition by the analysis of application class tree the marker levels in experimenter's the sample and the marker levels in the reference sample are compared.Classification tree analysis can for example be finished by computer procedures.

On the other hand, this instructions provide a kind of for for give ABT-869 to the forecasting power for the treatment of cancer will be one or more each suffer from or doubt the methods that cancered experimenter classifies, the method is included in from measuring at least one in each experimenter's the sample and is selected from the level of following mark: NSE, CYFRA 21-1, CA125 and CEA, wherein following any indication cancer subtend experimenter gives the susceptibility of ABT-869: the level of NSE reduces with respect to the level of the NSE in the reference sample, the level of CA125 reduces with respect to the level of the CA125 in the reference sample, the level of CYFRA 21-1 reduces with respect to the level of the CYFRA 21-1 in the reference sample, the level of CEA raises with respect to the level of the CEA in the reference sample, or its any combination.The method can comprise in addition based on the level of at least one among NSE, CA125, CYFRA 21-1 and the CEA each experimenter is categorized as responsive with ABT-869 treatment.In the method, one or more experimenters can suffer from or suspicious trouble non-small cell lung cancer.According to the method, for example the NSE level in experimenter's the sample can reduce with respect to the NSE level in the reference sample.CA125 level in experimenter's the sample can reduce with respect to the CA125 level in the reference sample.CYFRA 21-1 level in experimenter's the sample can reduce with respect to the CYFRA 21-1 level in the reference sample.CEA level in experimenter's the sample can raise with respect to the CEA level in the reference sample.The method can comprise the mark signature that produces each experimenter from the level of one or more marks in addition, wherein have the mark signature indication of preassigned pattern with respect to the experimenter with the mark signature that does not have preassigned pattern, the experimenter increases the susceptibility that gives ABT-869.The method can comprise in addition by application class tree analyzes level comparison with the identical mark in the marker levels in each experimenter's the sample and the reference sample, and classification tree analysis can be finished by computer procedures.In any above method, sample can be blood sample, comprises serum or plasma sample.Any above method can comprise the step that obtains sample from the experimenter in addition.In any above method, the level of each mark can for example be measured by immunohistochemistry or immunoassays.

On the other hand, this instructions provides a kind of cancer subtend experimenter for the prediction experimenter to give the kit of the susceptibility of ABT-869, the method comprises: the array that a) contains one or more binding reagents, each binding reagents is selected from following mark at least one and has independently binding specificity: NSE, CA125, CYFRA 21-1 and CEA, and wherein each binding reagents independently is combined with the discrete location on the matrix at least one by one; And b) contain the control sample of one or more marks of predeterminated level in the array, wherein the predeterminated level of each mark is such level: this mark with respect to the level indication experimenter's of this level cancer to giving the susceptibility of ABT-869.The configuration kit can be non-small cell lung cancer with the cancer that prediction gives the susceptibility of ABT-869.In this kit, the NSE level in the control sample can be for example such level: the level of the NSE in experimenter's the sample is lower than this level, is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869.CA125 level in the control sample can be such level: the level of the CA125 in experimenter's the sample is lower than this level, is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869.CYFRA 21-1 level in the control sample can be such level: the level of the CYFRA 21-1 in experimenter's the sample is lower than this level, is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869.CEA level in the control sample can be such level: the level of the CEA in experimenter's the sample is higher than this level, is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869.In this kit, one or more matrix can respectively comprise the solid support with the coupling of detection property mark.This detection mark can comprise for example fluorescent chemicals.This kit can comprise in addition for the instructions of mensuration from each marker levels of experimenter's sample.Experimenter's sample can be blood sample, comprises plasma sample or blood serum sample.

On the other hand, this instructions provides a kind of cancer subtend experimenter for the prediction experimenter to give the kit of the susceptibility of ABT-869, it comprises: a) comprise the microarray that is selected from following one or more mark: NSE, CA125, CYFRA 21-1, CEA and clipped form thereof, and b) contain the control sample of one or more marks of predeterminated level, wherein the predeterminated level of each mark is such level: this mark with respect to the level indication experimenter's of this level cancer to giving the susceptibility of ABT-869.The configuration kit can be non-small cell lung cancer with the cancer that prediction gives the susceptibility of ABT-869.In this kit, the NSE level in the control sample can be for example such level: the level of the NSE in experimenter's the sample is lower than this level, is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869.CA125 level in the control sample can be such level: the level of the CA125 in experimenter's the sample is lower than this level, is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869.CYFRA 21-1 level in the control sample can be such level: the level of the CYFRA 21-1 in experimenter's the sample is lower than this level, is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869.CEA level in the control sample can be such level: the level of the CEA in experimenter's the sample is higher than this level, is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869.In this kit, one or more matrix can respectively comprise the solid support with the coupling of detection property mark.This detection mark can comprise for example fluorescent chemicals.This kit can comprise in addition for the instructions of mensuration from each marker levels of experimenter's sample.Experimenter's sample can be blood sample, comprises plasma sample or blood serum sample.

The accompanying drawing summary

Fig. 1 be show with or without three of the ABT-869 treatment on the same group 3/4 phase NSCLC patient's the Kaplan Meier figure of the overall survival in fate (OS) not.

Fig. 2 be one group with Alimta with or not with the Kaplan-Meier figure of patient's group (M05-780) of ABT-751 treatment, according to the baseline blood plasma level of the mark of comparing with NSCLC median threshold value, each of eight marks estimating is drawn OS.

Fig. 3 shows that two Kaplan Meier based on patient group's (" group 2 ") analysis scheme, and this patient group is characterised in that with the group patient who treats without ABT-869 and compares with OS increase after the ABT-869 treatment.

Describe in detail

A. Definition

The section header that uses in the whole instructions of this section and this paper is not intended to restriction.

Specify unless a) context is clear, otherwise singulative used herein comprises plural indicant.For the narration of this paper numerical range, expect that clearly each intervention between this scope with same accuracy is digital.For example, for scope 6-9, except 6 and 9, also expection numeral 7 and 8, and for scope 6.0-7.0, clearly expection numeral 6.0,6.1,6.2,6.3,6.4,6.5,6.6,6.7,6.8,6.9 and 7.0.

B) Neuronspecific enolase (" NSE ")

The term of the commutative use of this paper " neuronspecific enolase " and " NSE " refer to, by albumen and the conservative variant thereof of the people's gene coding that also is called as enolase 2 (the symbol EN02 of official).Term used herein " official's symbol " refers to the title used in the EntrezGene database of being safeguarded by American National biotechnology information center.

C) Cancer antigen 125 (" CA125 ")

The term of the commutative use of this paper " cancer antigen 125 " and " CA125 " refer to, be identified as the tumor markers of oophoroma and derive from the Carbohydrate Antigens of the albumen of the relevant MUC-1 6 (also being called as MUC16) of cell surface (its for by people MUC16 gene (the symbol M UC16 of official)) coding, and the conservative variant of CA125.

D) The serum soluble fragment of Cyfra21-1 (" CYFRA 21-1 ")

The term of the commutative use of this paper " the serum soluble fragment of Cyfra21-1 " and " CYFRA 21-1 " refer to, be identified as comprising lung cancer kinds cancer tumor markers and derive from the antigen of Cyfra21-1, Cyfra21-1 is the albumen by the conservative variant coding of human keratin 19 genes (the symbol KRT19 of official) and KRT19.

E) Carcinomebryonic antigen (" CEA ")

The term of the commutative use of this paper " carcinomebryonic antigen " and " CEA " refer to have people's albumen and the conservative variant thereof of the amino acid sequence under the GenBank accession number CAE75559.

F) Detection property mark

Term used herein " detect property mark " refers to, but produces any part of measuring-signal via the state variation of one or more molecules of light, electricity or the indication of other physics and this part coupling.Such physics indicant is contained spectroscopy, photochemistry, biological chemistry, immunochemistry, electromagnetism, radiochemistry and chemical method, such as but not limited to fluorescence, chemiluminescence, chemiluminescence etc.

G) The experimenter

Commutative use term used herein " experimenter " and " patient ", and no matter the experimenter has or standing any type for the treatment of.Term used herein " experimenter " refers to any vertebrate, include but not limited to mammal (for example, ox, pig, camel, yamma, horse, goat, rabbit, sheep, hamster, cavy, cat, dog, rat and mouse, inhuman primate (such as monkey (such as machin (cynomolgous monkey)), chimpanzee etc.) and people).Preferably, the experimenter is the people.

H) Specimen

Term used herein " specimen " refers generally to, the tested and/or doubtful biomaterial that contains one or more cancer markers.Biomaterial can derive from any biogenetic derivation.The example of biomaterial includes but not limited to the peripheral blood sample, tumour or doubtful tumor tissues, the thin-layer cell product that imitate, the Fine needle aspiration sample, the marrow sample, the lymph node sample, urine sample, the ascites sample, the lavation sample, oesophagus grooming sample, bladder or lung washing sample, the spinal fluid sample, brain liquid sample, conduit is drawn sample, the nipple discharge sample, the pleural effusion sample, FF tissue sample, paraffin-embedded tissue sample or the extract that from any peripheral blood sample, produces or the sample of processing, the serum of blood sample or blood plasma part.Specimen can directly be used after available from biogenetic derivation, or uses after pre-service is with the feature that changes sample.For example, such pre-service can comprise from blood and prepares blood plasma, dilution viscous liquid etc.Pretreated method also can relate to filtration, precipitation, dilution, distillation, mixing, concentrated, deactivation interfering component, add reagent, cracking etc.If use such preprocess method with respect to specimen, so such preprocess method so that cancer cell be retained in the specimen.

B. The prediction cancer is to the mark of the susceptibility of ABT-869

Method of the present disclosure and kit part is based on unexpected following the discovery: the level of the special sign thing that exists in the specimen available from the experimenter (or " biomarker ") is that experimenter's cancer is to the prediction of the susceptibility that gives ABT-869.These predictability marks comprise NSE, CA125, CYFRA 21-1 and CEA.

Method of the present invention is particularly useful for compd A BT-869 (Linifanib; [N-(4-(3-amino-l H-indazole-4-yl) phenyl)-N'-(2-fluoro-5-aminomethyl phenyl) urea]), it is competitive receptor tyrosine kinase (RTK) inhibitor of ATP, and it is growth factor (PDGF) the receptor family member's in vascular endothelial growth factor (VEGF) and blood platelet source strong inhibitor.Referring to Shankar D.B. etc., Blood, April 15: 109 (8), 3400-8 (2007).The chemical constitution of ABT-869 is:

Figure 201180033733X100002DEST_PATH_IMAGE002

Described ABT-869 other synthetic methods (referring to, such as A. Kruger etc., Org. Process Res. Dev.13 (6), 1419-25 (2009)).The method of administration and the method that comprise the pharmaceutical composition of ABT-869 and be used for treatment of cancer are known, and be specified in for example u.s. patent application serial number 11/636,189 (US 2007/0135387), its whole instructions is incorporated herein by reference.

The predictability mark is any mark that can exist and measure in from experimenter's specimen (for example can be the blood sample of blood plasma or blood serum sample), and the level of this mark in the sample (i.e. amount) is closed particular treatment compound and/or other reacting phase of compounds with cancer.Found that mark NSE as herein described, CA125, CYFRA 21-1 and CEA are the experimenters, or more particularly, experimenter's cancer means to give ABT-869 to the prediction with the susceptibility of ABT-869 treatment with the ABT-869 treatment.In order to measure mark and clinical effectiveness, more specifically with correlativity to the susceptibility of ABT-869, measured the mark concentration among the experimenter who suffers from the specific objective cancer, for example for base measurement at the zero-time point measurement, then after the begin treatment scheme, at the second point in time measurement of the 21st or 22 day for example of about 3 weeks.Mark threshold value or " cutoff (cut-off) " can for example be established as median for the particular cancers type, or can select any other statistical method of such cutoff to set up in value distributes by using.For each mark, the experimenter is categorized as has the marker levels that is higher or lower than this threshold value.Then measure survival (for example overall survival) as the function for the treatment of classification, and compare for each mark and treatment.

Therefore, for each mark, the reference levels of identifying predetermined cutoff level and providing thereby can use according to method as herein described and kit.More specifically, as described in this paper other places, the level that the level that the level that the level of NSE is lower than predeterminated level, the CYFRA 21-1 of NSE is lower than the predeterminated level of CYFRA 21-1, CA125 is lower than the predeterminated level of CA125, CEA is higher than predeterminated level or its any combination of CEA, indication be higher than with respect to the level of NSE, CA125 or CYFRA 21-1 each mark predeterminated level the experimenter or be lower than the experimenter of the predeterminated level of CEA with respect to the level of CEA, experimenter's cancer increases the susceptibility that gives ABT-869.

Common use immunohistochemistry or immunoassay (for example enzyme immunoassay (EIA) (EIA), and its kit is from the easy commercially available acquisition of many commercial supplier) measure from each marker levels in experimenter's the specimen.A kind of exemplary particulate enzyme immunoassay technique is the AXSYM system available from Abbott Laboratories.This mensuration can relate to multiple technology, in order to can measure the level of two or more marks from the output of single mensuration process.Any two or more marker levels of NSE, CA125, CYFRA 21-1 and CEA in the specimen capable of being combined, to produce mark signature (being sometimes referred to as " biomarker overview "), it is characterized in that the pattern that is formed by two or more marker levels at least.A kind of exemplary this pattern is lower than NSE by the level of for example NSE predetermined cutoff value forms together with following one or more: the level that the level that the level of CA125 is lower than the predetermined cutoff value of CA125, CYFRA 21-1 is lower than the predetermined cutoff value of CYFRA 21-1 and CEA is higher than the predetermined cutoff value of CEA.The mark signature can comprise the level of the one or more marks except NSE, CA125, CYFRA 21-1 and CEA.Have the mark signature indication of preassigned pattern (namely satisfying specific criteria, for example each cutoff standard of at least two marks) with respect to the mark signature that does not have preassigned pattern, the experimenter increases the susceptibility that gives ABT-869.

The basis of in the method for this instructions and kit, using these marks to provide to use the treatment of ABT-869 exploitation target on cancer.These methods can be especially as the basis of following mensuration that for example is used for the ABT-869 treatment, the ABT-869 treatment gives the experimenter as single therapy or as the part with the combined therapy of other chemotherapy (for example conventional chemotherapy).For having determined that marker levels is cancer to any type of cancer of the prediction of the susceptibility that gives ABT-869, can carry out these methods.A kind of exemplary this cancer is any cancer, for example non-small cell lung cancer, or any solid tumor.

C. Method

The method that gives the susceptibility of ABT-869 for the cancer subtend experimenter who predicts the experimenter relates to: the level of measuring at least one predictability mark as herein described (being serum soluble fragment (CYFRA 21-1) and the carcinomebryonic antigen (CEA) of neuronspecific enolase (NSE), cancer antigen 125 (CA125), Cyfra21-1).Below any or multiple indication be higher than than the level of NSE, CA125 or CYFRA 21-1 each mark predeterminated level the experimenter or be lower than the experimenter of the predeterminated level of CEA than the level of CEA, experimenter's cancer increases the susceptibility that gives ABT-869: 1) level of NSE is lower than the predeterminated level of NSE, 2) level of CA125 is lower than the predeterminated level of CA125, and CYFRA 21-1 is lower than the predeterminated level and 3 of CYFRA 21-1) level of CEA is higher than predeterminated level or its any combination of CEA.These methods can for example comprise all four the levels among NSE, CA125, CYFRA 21-1 and the CEA of measuring.The related cancer of this instructions contains the anti-angiogenic generation of expection treats for example any cancer of ABT-869 treatment, and especially contain any solid tumor (comprising tumor of breast) and cancer (comprising hepatocellular carcinoma, clear-cell carcinoma, small cell carcinoma and large cell carcinoma) and combination thereof, and comprise for example non-small cell lung cancer (NSCLC).

Based on the medicine kill cancer cell or reduce tumor size and/or reduce the ability of the growth of overall cancer or diffusion (transfers), cancer or experimenter (patient) can be described as comprising that the selected medicine scheme that gives ABT-869 is responsive or resisting.Insensitive cancer cell or tumour are regarded as the therapeutic scheme opposing, and are to nonreactive those cancer cells of pharmaceutical admixtures or tumour, and for example wherein pharmaceutical admixtures can not significantly reduce tumor size or slow down tumor growth or those cancer cells or the tumour of diffusion.Cancer cell to the therapeutic scheme sensitivity is the cancer cell that really pharmaceutical admixtures is reacted, and causes that tumor size minimizing and/or tumor growth or diffusion slow down, and (" OS ") increases thereby also cause overall survival.To the monitoring of pharmaceutical admixtures reaction can pass through many pathology, clinical and formation method is realized, those of those and the medical domain common general knowledge described of this paper other places for example.For example, can use any soft-tissue imaging technology (for example ultrasonic, CT and/or DCE-MRI) to estimate tumor size.Should also be understood that these methods can comprise in addition that any sample of tissue technology of use (including but not limited to blood drawing and fingerstick) and biopsy technology (comprising needle puncture biopsy) obtain specimen from the experimenter.

When measuring the level of two or more marks, the method can comprise the mark signature that produces the experimenter from the level of two or more marks in addition.The mark signature can comprise for example two or marker levels, and wherein this mark limits the feature that mark is signed with respect to each level of cutoff level, and these features form this signature together.The signature indication of total preassigned pattern (i.e. reflection has the pattern with respect to the marker levels of the particular kind of relationship of the cutoff of each mark separately) is with respect to the mark signature that does not have preassigned pattern, and the experimenter increases the susceptibility that gives ABT-869.For example, the indication experimenter increases the susceptibility that gives ABT-869 and is to be characterised in that following pattern based on the pre-determined signature pattern of all marker levels of NSE, CA125, CYFRA 21-1 and CEA: 1) level of NSE is lower than the predeterminated level of NSE, 2) level of CYFRA 21-1 is lower than the predeterminated level of CYFRA 21-1,3) level of CA125 is lower than the predeterminated level of CA125, and 3) level of CEA is higher than the predeterminated level of CEA.Any signature with all these pattern features is that the indication experimenter is to the exemplary signature of the susceptibility that gives ABT-869.

The analysis of marker levels can comprise in addition that with the level of the identical mark in the level of at least two marks and the control sample relatively this can analyze by the application class tree and finish.Classification tree analysis is common general knowledge, and can use computer procedures that it easily is applied to the analysis of marker levels.For example, can produce reflection and cancer to the reference 3D profile diagram of the relevant marker levels as herein described of the susceptibility for the treatment of with ABT-869.For any given experimenter, can produce comparable 3D figure, and will scheme and relatively indicate the experimenter that the mark of the susceptibility that gives ABT-869 is signed to determine whether that the experimenter has with reference to 3D figure.Classification tree analysis is fit to analysis mark thing level very much, and figure shows and easily explanation because they are especially obeyed.Yet will understand, can use any computer based to use, it is relatively from two different experimenters or from a plurality of marker levels of reference sample and experimenter, and provides the indication experimenter output of the susceptibility to giving ABT-869 based on method as herein described.

These methods can be used for one or more each trouble or doubt cancered experimenter and classify, give the ABT-869 effect of the cancer to treating the experimenter for prediction.Such method is included in the level of measuring at least one mark among NSE, CA125, CYFRA 21-1 and the CEA in the sample from each experimenter, and with the level of each mark and its level in the reference sample relatively.Reference sample comprises the amount of each mark corresponding with the predetermined cutoff value of mark.Below any indication cancer subtend experimenter give the susceptibility of ABT-869: 1) level of NSE reduces with respect to the level of the NSE in the reference sample, 2) level of CYFRA 21-1 reduces with respect to the level of the CYFRA 21-1 in the reference sample, the level of CEA raises with respect to the level of the CEA in the reference sample, the level of CA125 reduces with respect to the level of the CA125 in the reference sample, or its any combination.Therefore, these methods for example can be used for ABT-869 treatment wherein compare with alternative treatment may produce more excellent result patient colony as target.

D. Kit

This instructions also is provided for predicting that experimenter's cancer subtend experimenter gives the kit of the susceptibility of ABT-869.Kit can comprise for example array of one or more binding reagents, with the control sample of the one or more marks that contain predeterminated level, wherein the predeterminated level of each mark is such level: this mark indicates experimenter's cancer to giving the susceptibility of ABT-869 with respect to the level of this level.The predeterminated level of each mark is for example according to for example this paper other places determined cutoff of (for example among the embodiment) described statistical study or threshold value.Each binding reagents has independently binding specificity among NSE, CA125, CYFRA 21-1 and the CEA at least one.Exemplary this binding reagents is antibody.Alternatively, kit can comprise the array of two or more marks or their clipped form or fragment.

Antibody

Binding reagents can be for example polyclonal antibody, monoclonal antibody, chimeric antibody, people's antibody, affinity maturation antibody or antibody fragment.Can use the sandwich immunoassay form, wherein trapping antibody and detection antibody both are used for each mark.Antibody can be combined (for example puting together) with detection property mark.Although monoclonal antibody to mark/antigen high special, can preferably be used as trapping antibody so that mark/antigen as much as possible is fixed with polyclonal antibody.Then can preferably the detection antibody of the monoclonal antibody of inherent higher binding specificity as each mark/antigen will be had to mark/antigen.Under any circumstance, capture and detect the non-overlapped epi-position on each mark of antibody recognition, preferably do not disturb another combination.

By immunogene injection (for example subcutaneous or intramuscular injection) is produced polyclonal antibody in suitable non-human mammal (for example mouse or rabbit).Usually, immunogene can be induced the antibody that target antigen is had relatively high affinity that produces high-titer.If need, mark can be puted together by conjugation techniques well known in the art and carrier protein.Carrier commonly used comprises keyhole limpet hemocyanin (KLH), thyroglobulin, bovine serum albumin(BSA) (BSA) and tetanus toxoid.Then conjugate is used for immune animal.Then from the blood sample of taking from animal, obtain antibody.For generation of the technology of polyclonal antibody extensively be described in the document (referring to, for example, Methods of Enzymology, " Production of Antisera with Small Doses of Immunogen:Multiple Intradermal Injections (producing antiserum with low dose of immunogene: repeatedly intracutaneous injection); " Langone, Deng chief editor (Acad. Press, 1981)).The polyclonal antibody that is produced by animal can be further purified, for example by with in conjunction with the matrix of target antigen in conjunction with and on this matrix wash-out.One skilled in the art will know that for the common multiple technologies of the field of immunology of purifying and/or concentrated polyclone and monoclonal antibody (referring to, for example, Coligan, Deng Unit (1991) the 9th, Current Protocols in Immunology, Wiley Interscience).

For many application, preferred monoclonal antibody (mAb).Conventional method for generation of the hybridoma of secreting mAb is known (Kohler and Milstein (1975) Nature, 256:495).In brief, as described in Kohler and Milstein, this Technology Need: separate lymphocyte (wherein sample is available from the surgery sample) from the cancer patient's of 5 independent trouble melanoma, teratocarcinoma, cervical carcinoma, glioma or lung cancer local drainage lymph node, merge cell and with cell and SHFP-1 fusion.Screening produces the hybridoma in conjunction with the antibody of cancerous cell line.Can use routine screening technology (for example enzyme linked immunosorbent assay (ELISA), or " ELISA ") to measure the fundamental reaction pattern of target mAb, realize specific conclusive evidence between the mAb.

The antibody fragment of conjugated antigen also contained in term used herein " antibody ", single-chain antibody (scFv or other) for example, and it can produce with display technique of bacteriophage/select.Express the ability of antibody fragment on the virus (bacteriophage or bacteriophage) of bacterial infection surface so that can make unijunction close antibody fragment to separate with for example library above 1010 non-binding clones.In order to express antibody fragment (phage display) on the surface of bacteriophage, the antibody fragment gene (for example is inserted into coding phage surface albumen, pIII) in the gene, antibody fragment-pIII fusion show at phage surface (McCafferty etc. (1990) Nature, 348:552-554; Hoogenboom etc. (1991) Nucleic Acids Res. 19:4133-4137).

Because the lip-deep antibody fragment of bacteriophage is functional, so can separate with non-binding bacteriophage by the bacteriophage that the antigen affinity chromatography will be carried the antibody fragment of conjugated antigen.(McCafferty etc. (1990) Nature, 348:552-554).The affinity that depends on antibody fragment, single-wheel affinity select to obtain 20 times to 1,000,000 times enrichment factor.By the phage-infect bacterium with dilution, yet, can cultivate more bacteriophage and make it stand another and take turns selection.By this way, in taking turns 1000 times are enriched in two-wheeled and become 1,000,000 times (McCafferty etc. (1990) Nature, 348:552-554) in selecting.Therefore, be low (Marks etc. (1991) J. Mol. Biol. 222:581-597) even work as enrichment, the affinity of many wheels select also can cause the separation of rare bacteriophage.Because select phage antibody library to cause enrichment for antigen, thus few to the selection of 3-4 wheel most clone's conjugated antigen.Therefore only need clone (hundreds of) to relatively small amount to analyze combination with antigen.

By showing that in bacteriophage very large and various V gene pool can produce people's antibody and not need formerly immunity (Marks etc. (1991) J. Mol. Biol. 222:581-597).In one embodiment, separate natural VH and the VL storehouse that exists the human peripheral lymphocyte by PCR from non-immune donor.Montage is together to produce the scFv gene pool at random with the V gene pool can to use PCR, and the scFv gene pool can be cloned in the phage vector to produce the library (the same) of 3,000 ten thousand phage antibodies.From list " naivety " phage antibody library, Separated pin is to surpassing 17 the not binding antibody fragment of synantigen (comprising haptens, polysaccharide and albumen) (Marks etc. (1991) J. Mol. Biol. 222:581-597; Marks etc. (1993). Bio/Technology. 10:779-783; Griffiths etc. (1993) EMBO J. 12:725-734; Clackson etc. (1991) Nature. 352:624-628).Produced for the oneself protein antibody of (comprising human thyroglobulin, immunoglobulin (Ig), TNF and CEA) (Griffiths etc. (1993) EMBO J. 12:725-734).The antigen high special of antibody fragment to being used for selecting, and affinity (Marks etc. (1991) the J. Mol. Biol. 222:581-597 with 1 nM-100 nM scope; Griffiths etc. (1993) EMBO J. 12:725-734).Larger phage antibody library causes more antigens to larger proportion to have the separation of the antibody of higher binding affinity.

One of ordinary skill in the art will readily recognize that also can be by arbitrary Dispersal risk in many commerce services (for example, Berkeley Antibody Laboratories, Bethyl Laboratories, Anawa, Eurogenetec etc.).

Solid phase

In the kit of this instructions, each binding reagents can be combined with solid phase.Solid phase can be has enough surface affinity with any suitable material of binding antibody, and antibody for example has each trapping antibody of specific binding to one of mark.Solid phase can be taked any in the various ways, for example magnetic-particle, pearl, test tube, microtiter plate, cup, film, support molecule, quartz crystal, film, filter paper, disk or chip.Useful solid phase material comprises: natural polymer carbohydrates and synthetic modification thereof, the crosslinked or derivant that replaces, for example agar, agarose, crosslinked alginic acid, replacement and crosslinked guar gum, the cellulose esters, mixed cellulose ester and the cellulose ether that especially form with nitric acid and carboxylic acid; The natural polymer that comprises nitrogen, for example albumen and derivant comprise gelatin crosslinked or that modify; Native hydrocarbon polymkeric substance, for example latex and rubber; Synthetic polymer, polyvinyl for example, the derivant that comprises tygon, polypropylene, polystyrene, Polyvinylchloride, polyvinyl acetate (PVA) and partial hydrolysis thereof, polyacrylamide, polymethacrylate, the multipolymer of above-mentioned condensed polymer and terpolymer, for example polyester, polyamide, and other polymkeric substance, for example polycarbamate or polyepoxide; Inorganic material, for example the sulfate of earth alkali metal and magnesium or carbonate comprise barium sulphate, calcium sulphate, calcium carbonate, the silicate of alkali and alkaline earth metal ions, aluminium and magnesium; With aluminium or Si oxide or hydrate, for example clay, aluminium oxide, talcum, porcelain earth, zeolite, silica gel or glass (these materials can be used as the filter of above-mentioned polymeric material); With potpourri or the multipolymer of above-mentioned classification, the graft copolymer that obtains of the polymerization by initial synthetic polymer on the natural polymer that is pre-stored in for example.All these materials can be suitable shape (for example film, thin slice, pipe, particle or plate) use, perhaps they can be coated with, in conjunction with or be laminated to suitable inert carrier, such as paper, glass, plastic sheeting, fabric etc.Cellulose nitrate has excellent absorption and characterization of adsorption to the plurality of reagents that comprises monoclonal antibody.Nylon also has similar characteristic, and also is suitable.Any above-mentioned material can be used for forming the array of one or more specificity combinating reagents, for example microarray.

Alternatively, solid phase can form particulate.The particulate that is used for this instructions can be selected from the particulate material of any adequate types by those skilled in the art, and comprises the particulate that is comprised of polystyrene, polymethacrylate, polypropylene, latex, teflon, polyacrylonitrile, polycarbonate or similar material.In addition, particulate can be magnetic or paramagnetic particles, in order to help the operation of particulate in magnetic field.In an exemplary embodiment, particulate is carboxylated magnetic particle.Particulate can be suspended in the potpourri of soluble reagents and specimen, maybe can be by support material maintenance and fixing.In the latter's situation, on the support material or in particulate can not move in fact other positions in the support material.Alternatively, can by the precipitation or centrifugal from the suspending liquid the potpourri of soluble reagents and specimen separating particles.When particulate is magnetic or paramagnetism, can be by magnetic field separating particles from suspending liquid the potpourri of soluble reagents and specimen.The method of this instructions comprises automatically and automanual system that applicable to the system that uses the particulate technology wherein solid phase comprises particulate.This system comprises and is described in unsettled U. S. application numbers 425,651 and U.S. Patent number 5,089,424 (it corresponds respectively to European application EP 0 425 633 and the EP 0 424 634 of announcement) and the U.S. Patent number 5,006,309 those.

Affect other considerations that solid phase selects and comprise, make mark entity the minimized ability of non-specific binding and with the compatibility of employed Mk system.For example, the solid phase of using with fluorescence labeling should have enough low background fluorescence to allow input.After connecting the specificity trapping antibody, can be in addition with the surface of material (for example serum, albumen or other sealers) processing solid support so that non-specific binding minimize.

Detection system

The kit of this instructions can comprise one or more detection marks.One or more specificity combinating reagents (for example antibody) can be combined with detection property mark.The detection mark that is fit to use comprises any compound or the composition with the part that can pass through spectroscopy, photochemistry, biological chemistry, immunochemistry, electricity, light or chemical method detection.Such mark comprises, for example enzyme, oligonucleotides, nano particle chemiluminescence group, fluorophore, fluorescence quencher, chemiluminescence quencher or biotin.Therefore, for example, be configured to use in the immunoassay kit of light signal, light signal be measured as the chemiluminescence, fluorescence, phosphorescence, electrochemiluminescence, uv absorption, visible absorbance, infrared absorption, refraction, the resonance of surperficial plasmon that depend on analyte concentration change.Being configured to use in the immunoassay kit of electric signal, is that electric current, resistance, electromotive force, mass-to-charge ratio or the Ion Counting that depends on analyte concentration changes with electric signal measurement.In the immunoassay kit of the variation that is configured to use status signal, be that size, solubleness, quality or the resonance that depends on analyte concentration changes with the measure of the change of status signal.

The useful mark of this instructions comprises magnetic bead (for example, Dynabeads ^TM), fluorescent dye (for example, fluorescein, Texas Red, rhodamine, green fluorescent protein) etc. (referring to, for example, Molecular Probes, Eugene, Oreg., USA), chemiluminescence compound (acridine (acridinium) (for example, acridine-9-formamide) for example, phenanthridines (phenanthridinium), dioxetane, luminol etc.), radioactive label (3H for example, 125I, 35S, 14C or 32P), catalyzer (enzyme (commonly used horseradish peroxidase in ELISA for example for example, alkaline phosphatase, and colorimetric mark (for example collaurum (for example the diameter dimension scope of efficient scattering green glow is the gold grain of 40-80 nm)) or coloured glass or plastics (polystyrene for example beta galactosidase etc.)), polypropylene, latex etc.) pearl.Instruct the patent of the use of such mark to comprise U.S. Patent number 3,817,837,3,850,752,3,939,350,3,996,345,4,277,437,4,275,149 and 4,366,241.

Mark can with biological sample contact before, during or is connected with each antibody in the sandwich immunoassay form afterwards, for example be connected with detection antibody.So-called " directly mark " is directly to be connected with antibody before using in mensuration or to be incorporated into detection mark in the antibody.Can by well known to a person skilled in the art in the several different methods any with direct mark with detect antibody and be connected or be incorporated in the detection antibody.

By contrast, so-called " indirect labelling " a certain moment between test period is combined with each antibody usually.Often, indirect labelling is combined with using the part that connects with detection material before or be incorporated in the detection material.Therefore, for example, can be with each antibody biotinylation before in mensuration, using.Between test period, the fluorophore that avidin is puted together can be in conjunction with the detection material of carrying biotin to provide easily detected mark.

In another example of indirect labelling, polypeptide (for example polypeptide A or polypeptide G) that can the specific binding constant region for immunoglobulin also can be used as the mark for detection of antibody.These polypeptide are normal components of streptococcus cell membrane.They show that to the constant region for immunoglobulin from multiple species strong non-immunogenic is reactive (substantially referring to Kronval, Deng (1973) J. Immunol., 111:1401-1406, and Akerstrom (1985) J. Immunol, 135:2589-2542).Therefore such polypeptide can be labeled and add and measure in the potpourri, therein they will with respectively capture and detect antibody and be combined, and be combined with autoantibody, mark all and the composite signal that is caused by the analyte that exists in the sample and autoantibody is provided.

Some marks can need to use other reagent to produce detectable signal.In ELISA, for example, enzyme labeling (for example beta galactosidase) will need to add substrate (for example X-gal) to produce detectable signal.In the immunoassay kit that is configured to use acridine compounds as direct mark, also can in kit, comprise alkaline solution and hydrogen peroxide source.

The test kit of this instructions preferably includes for the instructions of mensuration from each marker levels (for example by carrying out one or more immunoassays) of experimenter's sample.This instructions can comprise the instructions for the specimen of analyzing particular type (blood sample for example, or more specifically, blood serum sample or plasma sample) in addition.The included instructions of the kit of this instructions is attachable to wrappage or can be used as the packing embolus and is included.Although instructions is normally hand-written or printing material, they are not limited to this.Can store such instructions and any medium that they pass to the terminal user is expected by this instructions.Such medium includes but not limited to electron storage medium (such as disk, audiotape, tape cassete, chip), optical medium (such as CD ROM) etc.Term used herein " instructions " can comprise the address of the internet site of the book that furnishes an explanation.

E. The adaptation of methods of this instructions

Those skilled in the art can recognize easily, and the embodiment of biomarker as herein described, oligonucleotides, method, kit and relevant composition typical example expressivity is not intended to scope of the present invention is limited.Can carry out different substitutions and modifications and not deviate from scope and spirit of the present invention disclosed herein instructions, this will be very apparent to those skilled in the art.

All patents of mentioning in the instructions and publication show this instructions one of ordinary skill in the art's level.All patents and publication are incorporated herein by reference to as showing each independent publication specifically and the identical degree of combination by reference individually.

This instructions that this paper illustrative is described can be implemented in the presence of the not concrete disclosed any or multiple key element of this paper, one or more restrictions not aptly.Therefore, for example, in each situation of this paper, term " comprises ", " basically by ... form " and " by ... form " in any all one of available other two terms replacement.The term that has used and wording are used as illustrative and non-limiting term, be not intended to get rid of in the use of this term and wording any equivalent or its part of the feature that shows and describe, and it should be understood that multiple modification is possible in the scope of this instructions requirement.Therefore, should be appreciated that, although this instructions is specifically open with optional feature by preferred embodiment, but those skilled in the art can take the modifications and variations of concept disclosed herein, and such modifications and variations are considered in the scope of the present invention that is limited by additional claims.

Embodiment

By embodiment but the mode that does not limit, will provide now the embodiment of this instructions.

Embodiment 1: based on the mark of the data of crossing over a plurality of NSCLC tests of using different treatments and the correlativity of clinical effectiveness

Three patients that distinguished by therapeutic scheme for the overall survival evaluation organize.All patients are diagnosed as 3/4 phase NSCLC.In NSCLC test at baseline by immunoassays survey mark substrate concentration.NSCLC experimenter is assigned to one of following three groups: M05-780(N=83), wherein the experimenter accepts pemetrexed (Alimta is available from Eli Lilly and Company, Indianapolis, IN) with or not with ABT-751 ((N-[2-[(4-hydroxy phenyl) amino]-the 3-pyridine radicals]-the 4-methoxybenzenesulphoismide, available from Abbott Laboratories, Abbott Park, IL); M05-782(N=21), wherein the experimenter accept docetaxel with or not with ABT-751; With M06-880 (N=103), wherein the experimenter accepts only ABT-869.

The patient is categorized as has the marker levels that is higher or lower than the threshold value marker levels.For each mark and the survival for the treatment of comparison as classification function.By several different methods assessment mark threshold value, method includes but not limited to middle pH-value determination pH, in community, determine be used for prediction NSCLC and the optimal threshold of optimum tuberculosis, the statistical modeling of value, and when treating with ABT-869, relatively suffer from stable disease to the relative concentration of the mark among the patient of rapid progress.

Fig. 1 shows three not on the same group Kaplan Meier figure of the overall survival in fate (OS-DUR) of 3/4 phase NSCLC patient, with the result of red display M05-780, show the result of M05-782 and show the result of M06-880 with blueness with green.Fig. 2 be one group with Alimta with or not with the Kaplan-Meier figure of patient's group (M05-780) of ABT-751 treatment, according to the baseline blood plasma level of each mark of comparing with NSCLC median threshold value, to each draws OS in eight blood plasma marker things estimating.Table 2 is provided at among ABT-869 treatment or the patient with the ABT-751 treatment, the viewed original logo thing level of 7 (Cyfra21-1, NSE, CEA, SCC, ProGRP, CA 15-3 and CA125) and the general introduction of threshold value in 8 marks among Fig. 2.

Fig. 3 shows two Kaplan Meier figure, both based on the patient group's who is accredited as group 2 further analysis.As shown in Figure 3, group 2 patients be cross over the NSCLC test when with Alimta with or do not show with the ABT-869 treatment with the patient of ABT-751 treatment relatively the time after remarkable those patients of increase of OS.According to baseline blood plasma marker thing water-glass syndrome 2 patients, all show following one or more: the level that the level that the level that the level of NSE is lower than the threshold value of NSE, CYFRA 21-1 is lower than threshold value, the CA125 of CYFRA 21-1 is lower than the threshold value of CA125, CEA is higher than the threshold value of CEA.

Table 2:

Figure 201180033733X100002DEST_PATH_IMAGE004

。

Claims

1. one kind is used for predicting that experimenter's cancer subtend experimenter gives the method for the susceptibility of ABT-869, said method comprising the steps of:

In available from experimenter's sample, measure at least one and be selected from the level of following mark: neuronspecific enolase (NSE), cancer antigen 125 (CA125), CYFRA 21-1 and carcinomebryonic antigen (CEA), wherein following any indication is with respect to NSE, the level of CA125 or CYFRA 21-1 be higher than each mark predeterminated level the experimenter or be lower than the experimenter of the predeterminated level of each mark with respect to the level of CEA, experimenter's cancer increases the susceptibility that gives ABT-869: the level of NSE is lower than the predeterminated level of NSE, the level of CYFRA 21-1 is lower than the predeterminated level of CYFRA 21-1, the level of CA125 is lower than the predeterminated level of CA125, the level of CEA is higher than predeterminated level or its any combination of CEA.

2. the process of claim 1 wherein that described cancer is non-small cell lung cancer.

3. the process of claim 1 wherein that described sample is blood sample.

4. the process of claim 1 wherein that described sample is serum or plasma sample.

5. the process of claim 1 wherein that described method comprises in addition from the experimenter obtains sample.

6. the process of claim 1 wherein that the level of described each mark measures by immunohistochemistry or immunoassays.

7. the method for claim 1, it comprises that at least two of mensuration are selected from the level of following mark: NSE, CYFRA 21-1, CA125 and CEA.

8. the method for claim 1, it comprises the level of measuring NSE, CA125, CYFRA 21-1 and CEA.

9. claim 7 or 8 method, wherein said method comprises the mark signature that produces the experimenter from the level of two or more marks in addition, wherein have the mark signature indication of preassigned pattern with respect to the mark signature that does not have preassigned pattern, the experimenter increases the susceptibility that gives ABT-869.

10. claim 7 or 8 method, wherein said method comprise in addition by the application class tree analyzes level comparison with the identical mark in the level of two or more marks in the sample and the control sample.

11. the method for claim 10, wherein said classification tree analysis is finished by computer procedures.

12. a cancer of predicting the experimenter said method comprising the steps of the method for the susceptibility that gives ABT-869:

In available from experimenter's sample, measure the marker levels in the mark group that comprises NSE, CA125, CYFRA 21-1 and CEA, and with the predeterminated level of each marker levels in the sample and each mark relatively, wherein each mark in the sample gives the susceptibility of ABT-869 with respect to the level indication cancer subtend experimenter of the predeterminated level of each mark.

13. the method for claim 12, wherein each marker levels in the sample and the predeterminated level of each mark are relatively comprised: with each marker levels in marker levels and the reference sample relatively, wherein said reference sample comprises each mark of the level corresponding with the predeterminated level of each mark.

14. the method for claim 12, wherein said cancer is non-small cell lung cancer.

15. the method for claim 12, wherein the NSE level in experimenter's the sample is lower than the predeterminated level of NSE.

16. the method for claim 12, wherein the CYFRA 21-1 level in experimenter's the sample is lower than the predeterminated level of CYFRA 21-1.

17. the method for claim 12, wherein the CEA level in experimenter's the sample is higher than the predeterminated level of CEA.

18. the method for claim 12, wherein the CA125 level in experimenter's the sample is lower than the predeterminated level of CA125.

19. the method for claim 12, wherein the sample from the experimenter is blood sample.

20. the method for claim 12, wherein the sample from the experimenter is serum or plasma sample.

21. comprising in addition from the experimenter, the method for claim 12, wherein said method obtain sample.

22. the method for claim 12, wherein the level of each mark in experimenter's the sample is measured by immunohistochemistry or immunoassays.

23. the method for claim 12, wherein said method comprises the mark signature that produces the experimenter from the level of mark in addition, wherein have the mark signature indication of preassigned pattern with respect to the experimenter with the mark signature that does not have preassigned pattern, the experimenter increases the susceptibility that gives ABT-869.

24. the method for claim 12, wherein said method comprise in addition by the analysis of application class tree the marker levels in experimenter's the sample and the marker levels in the reference sample are compared.

25. the method for claim 24, wherein said classification tree analysis is finished by computer procedures.

26. one kind be used for for give ABT-869 to the forecasting power for the treatment of cancer will be one or more each suffer from or doubt the methods that cancered experimenter classifies, described method is included in from measuring at least one in each experimenter's the sample and is selected from the level of following mark: NSE, CA125, CYFRA 21-1 and CEA, wherein following any indication cancer subtend experimenter gives the susceptibility of ABT-869: the level of NSE reduces with respect to the level of the NSE in the reference sample, the level of CYFRA 21-1 reduces with respect to the level of the CYFRA 21-1 in the reference sample, the level of CEA raises with respect to the level of the CEA in the reference sample, the level of CA125 reduces with respect to the level of the CA125 in the reference sample, or its any combination.

27. the method for claim 26, wherein said method comprise in addition based on the level of at least one among NSE, CYFRA 21-1, CA125 and the CEA each experimenter is categorized as responsive with the ABT-869 treatment.

28. the method for claim 26, wherein said one or more experimenters suffer from or doubt and suffer from non-small cell lung cancer.

29. the method for claim 26, wherein with respect to the NSE level in the reference sample, the NSE level in experimenter's the sample reduces.

30. the method for claim 26, wherein with respect to the CYFRA 21-1 level in the reference sample, the CYFRA 21-1 level in experimenter's the sample reduces.

31. the method for claim 26, wherein with respect to the CEA level in the reference sample, the CEA level in experimenter's the sample raises.

32. the method for claim 26, wherein with respect to the CA125 level in the reference sample, the CA125 level in experimenter's the sample reduces.

33. the method for claim 26, wherein said sample is blood sample.

34. the method for claim 26, wherein said sample are serum or plasma sample.

35. comprising in addition from each experimenter, the method for claim 26, wherein said method obtain sample.

36. the method for claim 26, the level of wherein said each mark is measured by immunohistochemistry or immunoassays.

37. the method for claim 26, wherein said method comprises the mark signature that produces each experimenter from the level of one or more marks in addition, wherein have the mark signature indication of preassigned pattern with respect to the experimenter with the mark signature that does not have preassigned pattern, the experimenter increases the susceptibility that gives ABT-869.

38. comprising in addition by application class tree, the method for claim 26, wherein said method analyze level comparison with the identical mark in the marker levels in each experimenter's the sample and the reference sample.

39. one kind is used for predicting that experimenter's cancer subtend experimenter gives the kit of the susceptibility of ABT-869, it comprises:

A. the array that contains one or more binding reagents, each binding reagents is selected from following mark at least one and has independently binding specificity: NSE, CYFRA 21-1, CA125 and CEA, and wherein the discrete location of each binding reagents at least one matrix independently is combined; With

B. the control sample that contains one or more marks of predeterminated level in the array, wherein the predeterminated level of each mark is such level: this mark indicates experimenter's cancer to giving the susceptibility of ABT-869 with respect to the level of this level.

40. the kit of claim 39, wherein said cancer is non-small cell lung cancer.

41. the kit of claim 39, wherein the NSE level in the control sample is such level: it is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869 that the level of the NSE in experimenter's the sample is lower than this level.

42. the kit of claim 39, wherein the CYFRA 21-1 level in the control sample is such level: it is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869 that the level of the CYFRA 21-1 in experimenter's the sample is lower than this level.

43. the kit of claim 39, wherein the CA125 level in the control sample is such level: it is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869 that the level of the CA125 in experimenter's the sample is lower than this level.

44. the kit of claim 39, wherein the CEA level in the control sample is such level: it is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869 that the level of the CEA in experimenter's the sample is higher than this level.

45. the kit of claim 39, wherein said one or more matrix respectively comprise the solid support with the coupling of detection property mark.

46. the kit of claim 45, wherein said detection mark comprises fluorescent chemicals.

47. the kit of claim 39, it comprises in addition for the instructions of mensuration from each marker levels of experimenter's sample.

48. the kit of claim 47, wherein the sample from the experimenter is blood sample.

49. the kit of claim 47, wherein the sample from the experimenter is plasma sample.

50. the kit of claim 47, wherein the sample from the experimenter is blood serum sample.

51. one kind is used for predicting that experimenter's cancer subtend experimenter gives the kit of the susceptibility of ABT-869, it comprises:

A. comprise the microarray that is selected from following one or more mark: NSE, CYFRA 21-1, CA125, CEA and clipped form thereof, and

B. the control sample that contains one or more marks of predeterminated level, wherein the predeterminated level of each mark is such level: this mark indicates experimenter's cancer to giving the susceptibility of ABT-869 with respect to the level of this level.

52. the kit of claim 51, wherein said cancer is non-small cell lung cancer.

53. the kit of claim 51, wherein the NSE level in the control sample is such level: it is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869 that the level of the NSE in experimenter's the sample is lower than this level.

54. the kit of claim 51, wherein the CYFRA 21-1 level in the control sample is such level: it is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869 that the level of the CYFRA 21-1 in experimenter's the sample is lower than this level.

55. the kit of claim 51, wherein the CA125 level in the control sample is such level: it is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869 that the level of the CA125 in experimenter's the sample is lower than this level.

56. the kit of claim 51, wherein the CEA level in the control sample is such level: it is that experimenter's cancer is to the indication of the susceptibility that gives ABT-869 that the level of the CEA in experimenter's the sample is higher than this level.

57. the kit of claim 51, it comprises in addition for the instructions of mensuration from each marker levels of experimenter's sample.

58. the kit of claim 51, wherein the sample from the experimenter is blood sample.

59. the kit of claim 51, wherein the sample from the experimenter is plasma sample.

60. the kit of claim 51, wherein the sample from the experimenter is blood serum sample.