JP5130999B2 - Anticancer drug efficacy prediction method - Google Patents
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本発明は 乳癌治療におけるトラスツズマブの奏功性ならびに無増悪期間の延長性を、トラスツズマブ投与前に予測する方法等に関するものである。 The present invention relates to a method for predicting the success of trastuzumab and the prolongation of progression-free period in the treatment of breast cancer before trastuzumab administration.
乳癌は、女性の間では最も発症率が高い悪性腫瘍の一つであり、５０〜５５歳の年齢層の女性では第１位の死因となっている。特に、受容体型チロシンキナーゼ（ＲＴＫ）をコードする遺伝子ＨＥＲ−２／ｎｅｕの過剰発現を示す乳癌は、臨床的に予後不良であることが知られている（非特許文献１）。また、このようなＨＥＲ−２／ｎｅｕ遺伝子の過剰発現は、原発性乳房腫瘍の２０〜２５％において認められることが報告されている（非特許文献２）。 Breast cancer is one of the most common malignant tumors among women, and is the leading cause of death among women aged 50 to 55 years. In particular, it is known that breast cancer showing overexpression of the gene HER-2 / neu encoding a receptor tyrosine kinase (RTK) has a clinically poor prognosis (Non-patent Document 1). Moreover, it has been reported that such overexpression of the HER-2 / neu gene is observed in 20 to 25% of primary breast tumors (Non-patent Document 2).
トラスツズマブ（trastuzumab；HERCEPTIN（登録商標））は、ＨＥＲ−２／ｎｅｕタンパクを標的とした、マウスＨＥＲ−２抗体４Ｄ５組換えヒト化抗体であり（特許文献１）、ＨＥＲ−２／ｎｅｕ過剰発現転移性乳癌患者の治療に有効であることが臨床的に明らかにされている（非特許文献３）。また、トラスツズマブは、種々の化学療法剤とも併用して治療に用いられており、組み合わせて使用される化学療法剤としては、パクリタキセル（非特許文献４及び５）、ドセタキセル（非特許文献６及び７）等のタキソイドや、アロマターゼインヒビター（非特許文献８）や、抗エストロゲン剤（非特許文献８及び９）や、リポソーム性ドキソルビシン（非特許文献１０）等が挙げられる。 Trastuzumab (HERCEPTIN (registered trademark)) is a mouse HER-2 antibody 4D5 recombinant humanized antibody targeting the HER-2 / neu protein (Patent Document 1), and HER-2 / neu overexpression transfer It has been clinically clarified that it is effective in the treatment of patients with metastatic breast cancer (Non-patent Document 3). Trastuzumab is also used for treatment in combination with various chemotherapeutic agents. Examples of chemotherapeutic agents used in combination include paclitaxel (Non-Patent Documents 4 and 5) and docetaxel (Non-Patent Documents 6 and 7). ), Aromatase inhibitors (Non-Patent Document 8), anti-estrogens (Non-Patent Documents 8 and 9), liposomal doxorubicin (Non-Patent Document 10), and the like.
最近になって、トラスツズマブによる治療を受けた患者のうち１０〜３０％に心毒性が認められることが臨床的に明らかにされた。このようなトラスツズマブの重篤な副作用を考慮すると、トラスツズマブ投与前に、トラスツズマブ治療の有効性をあらかじめ予測する方法を確立することが、乳癌の治療方針を立てる上で非常に重要である。しかし、他の抗癌剤と同様に、トラスツズマブの効果には個人差があり、効果が見られるかどうかは実際に治療を開始した後でなければ判定できないのが現状である。 Recently, it has been clinically demonstrated that 10-30% of patients treated with trastuzumab have cardiotoxicity. In view of such serious side effects of trastuzumab, it is very important to establish a method for predicting the efficacy of trastuzumab treatment before the administration of trastuzumab in advance of the treatment policy for breast cancer. However, as with other anticancer agents, there are individual differences in the effects of trastuzumab, and it can be determined only after actually starting treatment whether or not the effects are seen.
本発明の課題は、乳癌治療におけるトラスツズマブの奏功性ならびに無増悪期間の延長性を、トラスツズマブ投与前にあらかじめ予測する方法等を提供することにある。 An object of the present invention is to provide a method for predicting the success of trastuzumab in breast cancer treatment and prolonging the progression-free period in advance of the administration of trastuzumab.
本発明者らは、トラスツズマブ投与前の乳癌患者から採取した血漿中のＮ結合型糖鎖に関して質量分析を行い、３１のマススペクトルピークを検出した。これらの３１の糖鎖と、臨床情報との相関を検討した結果、トラスツズマブ療法奏功例（ｎｏｎ−ＰＤ群）において、２５３４ｍ／ｚの糖鎖の検出強度が、トラスツズマブ療法非奏功例（ＰＤ群）と比較して有意に高いこと、さらに、２５３４ｍ／ｚの糖鎖が高い群では、無増悪期間の有意な延長が認められることを確認した。以上の結果から、本発明者らは、２５３４ｍ／ｚ糖鎖の検出強度を指標として、乳癌治療におけるトラスツズマブの有効性の予測を行いうることを見い出し、本発明を完成した。 The present inventors performed mass spectrometry on N-linked sugar chains in plasma collected from breast cancer patients before trastuzumab administration, and detected 31 mass spectral peaks. As a result of examining the correlation between these 31 sugar chains and clinical information, the detection intensity of the sugar chain of 2534 m / z in trastuzumab therapy successful cases (non-PD group) was non-successful in trastuzumab therapy (PD group) In addition, it was confirmed that a significant increase in the progression-free period was observed in the group having a high sugar chain of 2534 m / z. Based on the above results, the present inventors have found that the efficacy of trastuzumab in breast cancer treatment can be predicted using the detection intensity of 2534 m / z sugar chain as an index, and the present invention has been completed.
すなわち本発明は、(１) トラスツズマブ（trastuzumab）投与前に乳癌治療におけるトラスツズマブの奏功性を分析するためのデータを収集する方法であって、癌患者より採取された血漿中の糖タンパクからＮ結合型糖鎖を遊離させる工程と、遊離させた糖鎖を精製する工程と、精製した糖鎖の質量分析を行う工程と、ＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機を用いたときの２５３４ｍ／ｚの糖鎖、又はそれに相当するピークの糖鎖の検出強度とトラスツズマブの奏功性とが、正の相関関係にあることを指標として、前記癌患者におけるトラスツズマブの奏功性を分析するためのデータを収集する工程とを、順次備えたことを特徴とする方法や、（２）トラスツズマブ（trastuzumab）投与前に乳癌治療におけるトラスツズマブによる無増悪期間の延長性を分析するためのデータを収集する方法であって、癌患者より採取された血漿中の糖タンパクからＮ結合型糖鎖を遊離させる工程と、遊離させた糖鎖を精製する工程と、精製した糖鎖の質量分析を行う工程と、ＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機を用いたときの２５３４ｍ／ｚの糖鎖、又はそれに相当するピークの糖鎖の検出強度とトラスツズマブの無増悪期間延長性とが、正の相関関係にあることを指標として、前記癌患者におけるトラスツズマブの無増悪期間の延長性を分析するためのデータを収集する工程とを、順次備えたことを特徴とする方法や、（３）トリプシン及びＮ−グリコシダーゼＦを用いて、Ｎ結合型糖鎖を遊離させることを特徴とする上記（１）又は（２）に記載の方法に関する。 That is, the present invention provides (1) trastuzumab (trastuzumab) A method of collecting data for analyzing successful of Torasutsuzuma blanking in breast cancer therapy prior to administration, N from glycoprotein in plasma taken from a cancer patient A step of releasing a conjugated sugar chain, a step of purifying the released sugar chain, a step of performing mass spectrometry of the purified sugar chain, and a 2534 m / z when using a MALDI-TOF-MS type analyzer. Collect data to analyze the efficacy of trastuzumab in the above cancer patients, using as an index the positive correlation between the detection intensity of the sugar chain or the peak sugar chain corresponding thereto and the response of trastuzumab and a step, and wherein, further comprising sequentially an extension of the time to progression by Torasutsuzuma Bed in breast cancer therapy prior to administration (2) trastuzumab (trastuzumab) A method of collecting data for analysis, and purifying the step of liberating the N-linked sugar chains from glycoproteins in the plasma taken from a cancer patient, the was released sugar chains were purified sugar The mass spectrometry of the chain, the detection intensity of the sugar chain of 2534 m / z when using a MALDI-TOF-MS type analyzer, or the peak sugar chain corresponding thereto, and the prolongation of trastuzumab without exacerbation period Collecting data for analyzing the prolongation of the progression-free period of trastuzumab in the cancer patient using the positive correlation as an index, and (3) The method according to (1) or (2) above, wherein the N-linked sugar chain is released using trypsin and N-glycosidase F.
本発明によると、乳癌患者から採取した血漿中の糖鎖を解析することにより、乳癌治療におけるトラスツズマブの奏功性や無増悪期間の延長性を、トラスツズマブ投与前にあらかじめ予測することが可能になる。 According to the present invention, by analyzing sugar chains in plasma collected from breast cancer patients, it is possible to predict in advance the efficacy of trastuzumab in breast cancer treatment and the prolongation of progression-free period before trastuzumab administration.
本発明は、乳癌治療におけるトラスツズマブの有効性（奏功性及び無増悪期間の延長性）を、トラスツズマブ投与前に予測する方法であって、（１）癌患者より採取された血漿中の糖タンパクからＮ結合型糖鎖を遊離させる工程と、（２）遊離させた糖鎖を精製する工程と、（３）精製した糖鎖の質量分析を行う工程と、（４）ＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機を用いたときは２５３４ｍ／ｚの糖鎖、ＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機以外の質量分析機を用いた場合は、ＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機を用いたときの２５３４ｍ／ｚに相当するピークの糖鎖、の検出強度とトラスツズマブの有効性とが、正の相関関係にあることを指標として、前記癌患者におけるトラスツズマブの有効性を予測する工程とを、順次備えたものであれば特に制限されず、より具体的には、例えばＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機を用いた質量分析により得られるマススペクトルの２５３４ｍ／ｚの糖鎖の検出強度と、予め決定した検出強度の閾値とを比較し、前記検出強度の測定値が前記閾値より高い場合にトラスツズマブが有効（奏功及び無増悪期間の延長）であると評価する、トラスツズマブの有効性予測方法を好適に例示することができる。上記トラスツズマブとは、マウスＨＥＲ−２抗体４Ｄ５組換えヒト化抗体であり、また、上記癌患者の有する癌は、乳癌であれば特に制限されず、再発または進行乳癌であっても、初期乳癌であってもよい。 The present invention relates to a method for predicting the effectiveness of trastuzumab in the treatment of breast cancer (response and prolongation of progression-free period) before trastuzumab administration, comprising: (1) from glycoproteins in plasma collected from cancer patients A step of releasing an N-linked sugar chain, (2) a step of purifying the released sugar chain, (3) a step of mass spectrometry of the purified sugar chain, and (4) MALDI-TOF-MS type analysis. 2534 m / z sugar chain when using the analyzer, and when using a mass analyzer other than the MALDI-TOF-MS analyzer, it corresponds to 2534 m / z when using the MALDI-TOF-MS analyzer And a step of predicting the efficacy of trastuzumab in the cancer patient by using, as an index, that the detection intensity of the sugar chain at the peak to be detected and the efficacy of trastuzumab are positively correlated. There is no particular limitation, and more specifically, for example, a detection intensity of a sugar chain of 2534 m / z in a mass spectrum obtained by mass spectrometry using a MALDI-TOF-MS type analyzer, and a threshold value of a predetermined detection intensity And a method for predicting the efficacy of trastuzumab, which evaluates that trastuzumab is effective (response and extension of the progression-free period) when the measured intensity value is higher than the threshold, can be suitably exemplified. The trastuzumab is a mouse HER-2 antibody 4D5 recombinant humanized antibody, and the cancer of the cancer patient is not particularly limited as long as it is breast cancer. There may be.
本発明において、Ｎ結合型糖鎖とは、糖タンパク質の糖鎖のうち、タンパク質のアスパラギン残基が持つ側鎖のアミド基の窒素原子に結合している糖鎖であり、Ｎ型糖鎖やアスパラギン結合型糖鎖とも称される。本発明において、血漿中の糖タンパクからＮ結合型糖鎖を遊離させる方法としては、例えば、Ｎ−グリコシダーゼＦ（グリコペプチダーゼ、ＰＮ Ｇａｓｅ、グリカナーゼ、グリコアミダーゼなどとも称される）やグリコペプチダーゼＡ等を用いた酵素法や、ヒドラジン分解法を具体的に例示することができるが、なかでも、Ｎ−グリコシダーゼＦによる酵素法を好例として挙げることができる。その際、トリプシン等のプロテアーゼを併用することもできる。また、本発明において、遊離させた糖鎖を精製する方法としては、試料中の混合物から糖鎖を選択的に捕捉し精製する方法であれば特に制限されないが、ＭＡＬＤＩ−ＴＯＦ−ＭＳでの高感度測定用に最適化された糖鎖補足ビーズであるBlotGlyco（登録商標）for MALDI（住友ベークライト株式会社製）を用いた方法を好例として具体的に挙げることができる。 In the present invention, the N-linked sugar chain is a sugar chain bonded to the nitrogen atom of the amide group of the side chain of the asparagine residue of the protein among the sugar chains of the glycoprotein. Also referred to as asparagine-linked sugar chain. In the present invention, methods for releasing N-linked sugar chains from glycoproteins in plasma include, for example, N-glycosidase F (also referred to as glycopeptidase, PNGase, glycanase, glycoamidase, etc.), glycopeptidase A, etc. Specific examples include an enzymatic method using hydrazine and a hydrazine decomposition method. Among them, an enzymatic method using N-glycosidase F can be cited as a good example. At that time, a protease such as trypsin can be used in combination. In the present invention, the method for purifying the released sugar chain is not particularly limited as long as it is a method for selectively capturing and purifying a sugar chain from a mixture in a sample, but it is not limited in MALDI-TOF-MS. A specific example is a method using BlotGlyco (registered trademark) for MALDI (manufactured by Sumitomo Bakelite Co., Ltd.), which is a sugar chain supplemented bead optimized for sensitivity measurement.
また、本明細書において、「ＭＡＬＤＩ−ＴＯＦ−ＭＳ」とは、Matrix Assisted Laser Desorption Ionization-Time-of-Flight（Mass Spectrometer）の略語である。ＭＡＬＤＩ法は、試料をプレート上にスポットした後、マトリクス溶液（2, 5-Dihydroxybenzoic acid）を添加、乾固し、結晶状態にし、パルスレーザー照射により大きなエネルギーをマトリクス上に与え、（Ｍ＋Ｈ）＋、（Ｍ＋Ｎａ）＋などの試料由来イオンとマトリクス由来イオンとを脱離させる方法で、ＭＡＬＤＩ−ＴＯＦ−ＭＳは、ＭＡＬＤＩ法を利用して飛行時間を元に質量を測定するものである。イオンが一定の加速電圧Ｖで加速される場合、イオンの質量をｍ、イオンの速度をｖ、イオンの電荷数をｚ、電気素量をｅ、イオンの飛行時間をｔとしたとき、イオンのｍ／ｚは、『ｍ／ｚ＝２ｅＶｔ２／Ｌ２』で表すことができる。本発明においては、ＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機以外の分析機を使用することもでき、イオン源として、例えば、電子イオン化法、化学イオン化法、電界離脱法、高速原子衝突法、エレクトロスプレーイオン化法、大気圧化学イオン化法等を用いることができ、また、分析法としては、例えば、磁場偏向型、四重極型、イオントラップ型、フーリエ変換イオンサイクロトロン共鳴型などの方法を用いることができる。さらに、上記分析法と、ＨＰＬＣとを組み合わせて用いることもできる。 Further, in this specification, “MALDI-TOF-MS” is an abbreviation of Matrix Assisted Laser Desorption Ionization-Time-of-Flight (Mass Spectrometer). In the MALDI method, after spotting a sample on a plate, a matrix solution (2,5-dihydroxybenzoic acid) is added, dried and crystallized, and a large amount of energy is given to the matrix by pulse laser irradiation. (M + H) + , (M + Na) + and other sample-derived ions and matrix-derived ions are desorbed, and MALDI-TOF-MS measures mass based on time of flight using the MALDI method. When an ion is accelerated at a constant acceleration voltage V, the ion mass is m, the ion velocity is v, the ion charge number is z, the elementary charge is e, and the ion flight time is t. m / z can be represented by “m / z = 2eVt 2 / L 2 ”. In the present invention, an analyzer other than the MALDI-TOF-MS type analyzer can also be used. As an ion source, for example, an electron ionization method, a chemical ionization method, an electric field separation method, a fast atom collision method, an electrospray ionization method. Method, atmospheric pressure chemical ionization method, and the like can be used, and as an analysis method, for example, a magnetic field deflection type, a quadrupole type, an ion trap type, a Fourier transform ion cyclotron resonance type, or the like can be used. . Further, the above analysis method and HPLC can be used in combination.
本発明において、ＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機を用いたときの２５３４ｍ／ｚの糖鎖とは、ＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機よる質量分析の結果、２５３４ｍ／ｚマススペクトルピークを呈する糖鎖であり、例えば、（Ｈｅｘ）５（ＨｅｘＮＡｃ）２（Ｄｅｏｘｙｈｅｘｏｓｅ）１＋（Ｍａｎ）３（ＧｌｃＮＡｃ）２や、（Ｈｅｘ）３（Ｄｅｏｘｙｈｅｘｏｓｅ）６ ＋（Ｍａｎ）３（ＧｌｃＮＡｃ）２等を具体的に挙げることができる。また、上記（Ｈｅｘ）５（ＨｅｘＮＡｃ）２（Ｄｅｏｘｙｈｅｘｏｓｅ）１＋（Ｍａｎ）３（ＧｌｃＮＡｃ）２や、（Ｈｅｘ）３（Ｄｅｏｘｙｈｅｘｏｓｅ）６ ＋（Ｍａｎ）３（ＧｌｃＮＡｃ）２等のＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機以外の質量分析機を用いた場合における、ＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機を用いたときの２５３４ｍ／ｚに相当するピークの糖鎖とは、ＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機を用いたときの２５３４ｍ／ｚマススペクトルピークを呈する糖鎖と相同の糖鎖を意味する。 In the present invention, a sugar chain of 2534 m / z when using a MALDI-TOF-MS type analyzer is a sugar chain that exhibits a 2534 m / z mass spectrum peak as a result of mass analysis by a MALDI-TOF-MS type analyzer. For example, (Hex) 5 (HexNAc) 2 (Deoxyhexose) 1+ (Man) 3 (GlcNAc) 2 and (Hex) 3 (Deoxyhexose) 6+ (Man) 3 (GlcNAc) 2 are specifically mentioned. be able to. Further, MALDI-TOF-MS type such as (Hex) 5 (HexNAc) 2 (Deoxyhexose) 1+ (Man) 3 (GlcNAc) 2 and (Hex) 3 (Deoxyhexose) 6+ (Man) 3 (GlcNAc) 2 When a mass spectrometer other than the analyzer is used, the peak sugar chain corresponding to 2534 m / z when the MALDI-TOF-MS type analyzer is used is the MALDI-TOF-MS type analyzer. It means a sugar chain that is homologous to the sugar chain that exhibits the 2534 m / z mass spectrum peak.
以下、実施例により本発明をより具体的に説明するが、本発明の技術的範囲はこれらの例示に限定されるものではない。 EXAMPLES Hereinafter, although an Example demonstrates this invention more concretely, the technical scope of this invention is not limited to these illustrations.
[乳癌患者からの検体の採取]
再発・進行乳癌患者に対しトラスツズマブ単独療法を受け、かつインフォームド・コンセントの得られた患者２４例について、治療開始前に血液を採取し、血漿を遠心分離した。採取した検体（血漿）は連結可能匿名化を行った後、−８０度で凍結保存した。
[血漿中の糖タンパク質糖鎖の分析]
糖タンパク質糖鎖の分析は、質量分析装置を用いて以下のように行った。２７μＬの血漿に、トリプシン及びＮ−グリコシダーゼＦを加えて反応させ、タンパク質と修飾糖鎖を遊離させた。遊離した糖鎖を、糖鎖捕捉ビーズを用いて選択的に捕捉し、ラベル化を行った。その後、ビーズに捕捉された糖鎖を精製、分離し、ＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機（Voyager-DETM STR Workstation；Applied Biosystems社製)により質量分析を行った。得られたシグナルは、内部標準と比較することにより定量化を行った。統計解析はｔ検定を用いた。また、質量分析の結果得られたシグナルから、GlycoSuite on line database（Proteome Systems）を用いて糖鎖の構造式を決定した。半数以上の症例で見られる３１種類の糖鎖を表１にまとめた。
[Collecting samples from breast cancer patients]
For 24 patients who received trastuzumab monotherapy for patients with relapsed / advanced breast cancer and obtained informed consent, blood was collected before starting treatment, and plasma was centrifuged. The collected specimen (plasma) was anonymized to be connectable and then stored frozen at -80 degrees.
[Analysis of glycoprotein sugar chains in plasma]
The analysis of glycoprotein sugar chains was performed as follows using a mass spectrometer. Trypsin and N-glycosidase F were added to and reacted with 27 μL of plasma to release the protein and modified sugar chain. The released sugar chain was selectively captured using a sugar chain capture bead and labeled. Thereafter, the sugar chain captured by the beads was purified and separated, and mass spectrometry was performed using a MALDI-TOF-MS type analyzer (Voyager-DE ™ STR Workstation; manufactured by Applied Biosystems). The signal obtained was quantified by comparison with an internal standard. Statistical analysis used t-test. Further, from the signal obtained as a result of mass spectrometry, the structural formula of the sugar chain was determined using GlycoSuite on line database (Proteome Systems). Table 1 shows 31 types of sugar chains found in more than half of the cases.
[糖鎖分析の結果解析]
得られた３１のピーク（糖鎖）について、臨床情報との相関を検討したところ、２５３４ｍ／ｚ糖鎖の検出強度はトラスツズマブ療法非奏功例(ＰＤ群；８例)では、４．３３±８．１３であるのに対し、トラスツズマブ療法奏功例（ｎｏｎ−ＰＤ群；１６例）では、１６．０７±１１．６０であり、ｎｏｎ−ＰＤ群において有意に高いことが明らかとなった (図１)。さらに、２５３４ｍ／ｚ糖鎖の検出強度の高い群(High group；１５例)は、低い群(Low group；９例)と比較して、無増悪期間の有意な延長が認められた(図２)。これらの結果から、血漿中の２５３４ｍ／ｚ糖鎖を指標として、トラスツズマブの治療効果の予測が可能であることが示された。
[Result analysis of glycan analysis]
The obtained 31 peaks (sugar chains) were examined for correlation with clinical information. The detected intensity of the 2534 m / z sugar chain was 4.33 ± 8 in the non-successful cases of trastuzumab therapy (PD group: 8 cases). In contrast, the number of patients successfully treated with trastuzumab (non-PD group; 16 cases) was 16.07 ± 11.60, which was significantly higher in the non-PD group (FIG. 1). ). Furthermore, the group with high detection intensity of the 2534 m / z sugar chain (High group; 15 cases) showed a significant prolongation of the progression-free period compared with the low group (Low group; 9 cases) (FIG. 2). ). From these results, it was shown that the therapeutic effect of trastuzumab can be predicted using 2534 m / z sugar chain in plasma as an index.
Claims (3)
（１）癌患者より採取された血漿中の糖タンパクからＮ結合型糖鎖を遊離させる工程；
（２）遊離させた糖鎖を精製する工程；
（３）精製した糖鎖の質量分析を行う工程；
（４）ＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機を用いたときの２５３４ｍ／ｚの糖鎖、又はそれに相当するピークの糖鎖の検出強度とトラスツズマブの奏功性とが、正の相関関係にあることを指標として、前記癌患者におけるトラスツズマブの奏功性を分析するためのデータを収集する工程； Trastuzumab (trastuzumab) A method of collecting data for analyzing successful of Torasutsuzuma blanking in breast cancer therapy prior to administration, wherein in that order with the following (1) to (4) Step ;
(1) releasing N-linked sugar chains from plasma glycoproteins collected from cancer patients;
(2) a step of purifying the released sugar chain;
(3) a step of performing mass spectrometry of the purified sugar chain;
(4) The detection intensity of the sugar chain of 2534 m / z when using a MALDI-TOF-MS type analyzer, or the peak sugar chain corresponding thereto, and the response of trastuzumab are positively correlated. Collecting data for analyzing the efficacy of trastuzumab in the cancer patient as an indicator;
（１）癌患者より採取された血漿中の糖タンパクからＮ結合型糖鎖を遊離させる工程；
（２）遊離させた糖鎖を精製する工程；
（３）精製した糖鎖の質量分析を行う工程；
（４）ＭＡＬＤＩ−ＴＯＦ−ＭＳ型分析機を用いたときの２５３４ｍ／ｚの糖鎖、又はそれに相当するピークの糖鎖の検出強度とトラスツズマブの無増悪期間延長性とが、正の相関関係にあることを指標として、前記癌患者におけるトラスツズマブの無増悪期間の延長性を分析するためのデータを収集する工程； Trastuzumab (trastuzumab) A method of collecting data for analyzing an extension of the time to progression by Torasutsuzuma Bed in breast cancer therapy prior to administration, the following (1) to (4) step sequence that comprises the A method characterized by:
(1) releasing N-linked sugar chains from plasma glycoproteins collected from cancer patients;
(2) a step of purifying the released sugar chain;
(3) a step of performing mass spectrometry of the purified sugar chain;
(4) The positive correlation between the detection intensity of the sugar chain of 2534 m / z when using a MALDI-TOF-MS type analyzer or the peak sugar chain corresponding to the sugar chain and the prolongation of trastuzumab without progression period Collecting data for analyzing the prolongation of the progression-free period of trastuzumab in the cancer patient as an index;
The method according to claim 1 or 2, wherein the N-linked sugar chain is liberated using trypsin and N-glycosidase F.
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