公开(公告)号：WO1994021821A1

公开(公告)日：1994-09-29

申请号：PCT/US1994002549

申请日：1994-03-15

Applicant: TROPIX, INC.

Inventor： TROPIX, INC. ,  BRONSTEIN, Irena, Y. ,  EDWARDS, Brooks ,  VOYTA, John, C.

IPC: C12Q01/68

CPC classification number: C07C43/1788 ,  C07C43/196 ,  C07C69/00 ,  C07C2603/00 ,  C07C2603/74 ,  C07D321/00 ,  C07F9/12 ,  C07F9/65512 ,  C07F9/65742 ,  C07H15/203 ,  C07H15/26 ,  C12Q1/34 ,  G01N21/76 ,  G01N33/581 ,  G01N33/582 ,  Y10S435/81

Abstract: Chemiluminescent bioassays for the presence or concentration of an analyte in a sample use 1,2-dioxetanes as substrates for the enzyme of an enzyme complex that bind to the analyte. The chemiluminescence obtained from the decomposition of the dioxetane trigerred by the enzyme through the formation of the corresponding 1,2-dioxetane oxyanion of the enzyme complex is enhanced by the addition of TBQ as an enhancement agent. Other polymeric quaternary onium salts can be used as enhancement agents in conjunction with enhancement additives which improve the ability of the enhancement agent to form hydrophobic regions in the aqueous sample, in which regions the 1,2-dioxetane oxyanion and its chemiluminescent decomposition products can be sequestered. A kit for performing such assays is also provided.

Abstract translation: 用于样品中分析物的存在或浓度的化学发光生物测定使用1,2-二氧杂环丁烷作为结合分析物的酶复合物的酶的底物。 通过添加TBQ作为增强剂，增强了通过酶复合物形成相应的1,2-二氧杂环丁烷氧阴离子而由酶分解的二氧杂环丁烷而得到的化学发光。 其他聚合季铵盐可用作增强剂与增强添加剂一起使用，增强添加剂改善增强剂在水样品中形成疏水区的能力，其中1,2-二氧环乙烷氧阴离子及其化学发光分解产物可以是 隔离。 还提供了用于进行这种测定的试剂盒。

公开(公告)号：WO1990002742A1

公开(公告)日：1990-03-22

申请号：PCT/US1989003836

申请日：1989-09-12

Applicant: TROPIX, INC.

Inventor： TROPIX, INC. ,  EDWARDS, Brooks ,  VOYTA, John, Charles

IPC: C07D321/00

CPC classification number: C07D321/00 ,  C07F9/65512

Abstract: Methods are disclosed for purifying chemiluminescent water-soluble 1,2-dioxetane derivatives suitable for use as reporter molecules in a variety of biological analytical systems, including enzyme-linked immunoassays, nucleic acid probe techniques, and structural determinations. The methods are based upon high pressure, medium pressure or low pressure liquid chromatography using as the stationary phase alkaline pH-stable compositions with the chromatographic characteristics of reversed-phase adsorbents, at alkaline pH values, and in the absence of acid-forming compounds or compounds with an unshared pair of electrons. Desalting of substantially pure water soluble 1,2-dioxetane derivatives may be accomplished by the same chromatographic systems or by molecular sieve chromatography systems, but in the absence of salt buffers. Under some circumstances, purification and desalting may be combined in a single chromatographic step. Substantially pure chemiluminescent water-soluble 1,2-dioxetane derivatives prepared by the methods of the invention are also disclosed.

公开(公告)号：WO1997014954A1

公开(公告)日：1997-04-24

申请号：PCT/US1996014389

申请日：1996-10-17

Applicant: TROPIX, INC.

Inventor： TROPIX, INC. ,  BRONSTEIN, Irena ,  EDWARDS, Brooks ,  SPARKS, Alison ,  VOYTA, John, C.

IPC: G01N21/76

CPC classification number: C12Q1/34 ,  C07D321/00 ,  C07F7/1804 ,  G01N33/52 ,  Y10S435/975

Abstract: Dioxetanes that can be triggered by bases are provided which can be used to detect bases in organic solvents, aqueous preparations and detect gaseous bases in the atmosphere. The dioxetanes can be used to detect the release of bases from various labels, as a means to detect the presence of a base released by physical or natural processes, to calibrate light measuring apparatus and to determine the amount of reducing or oxidizing agent present in the base.

Abstract translation: 提供可由碱触发的二氧杂环丁烷，可用于检测有机溶剂，含水制剂中的碱，并检测大气中的气态碱。 二氧杂环丁烷可用于检测各种标记物中碱的释放，作为检测通过物理或天然过程释放的碱的存在的手段，校准光测量装置并确定存在于其中的还原或氧化剂的量 基础。

公开(公告)号：WO1997014692A1

公开(公告)日：1997-04-24

申请号：PCT/US1996014390

申请日：1996-10-17

Applicant: TROPIX, INC.

Inventor： TROPIX, INC. ,  BRONSTEIN, Irena ,  EDWARDS, Brooks ,  SPARKS, Alison

IPC: C07D317/02

CPC classification number: G01N33/582 ,  C07D321/00 ,  C07F9/65512 ,  C07G3/00 ,  C12Q1/37 ,  Y02P20/55

Abstract: A new class of stable dioxetanes bears a polycyclic stabilizing group and aryloxy moiety, the oxygen atom of which is provided with a protective group which can be removed by an enzymatic or chemical trigger admixed with the dioxetane. The polycyclic stabilizing group is preferably spiroadamantane. The group further bears an alkoxy, aryloxy, aralkyloxy or cycloalkyloxy moiety which is partially or completely substituted with halogens, particularly fluorine and chlorine. Proper selection of electron active groups on the stabilizing moiety, the aryl group and the OR group yields enhanced enzyme kinetics, superior light intensity and excellent detection sensitivity in various assays.

Abstract translation: 一类稳定的二氧杂环丁烷具有多环稳定基团和芳氧基部分，其氧原子具有可以通过与二氧杂环丁烷混合的酶或化学触发剂除去的保护基团。 多环稳定基团优选为四金刚烷。 该基团还具有部分或完全被卤素，特别是氟和氯取代的烷氧基，芳氧基，芳烷氧基或环烷氧基部分。 稳定部分，芳基和OR基上的电子活性基团的适当选择在各种测定中产生增强的酶动力学，优异的光强度和优异的检测灵敏度。

公开(公告)号：WO1993013405A1

公开(公告)日：1993-07-08

申请号：PCT/US1992010803

申请日：1992-12-18

Applicant: TROPIX, INC.

Inventor： TROPIX, INC. ,  BRONSTEIN, Irena ,  EDWARDS, Brooks ,  VOYTA, John

IPC: G01N21/76

CPC classification number: G01N33/582 ,  G01N33/545 ,  Y10T428/31536 ,  Y10T428/31743 ,  Y10T428/31884

Abstract: A new and improved polymeric membrane for use in biological assays is provided. A blotting assay employing 1,2-dioxetanes as a source of chemiluminescent employs, as an improved membrane, a polymer comprised of at least one monomer of formula (1). The membranes reduce background signal, improve sensitivity and reliability.

Abstract translation: 提供了用于生物测定的新的和改进的聚合物膜。 使用1,2-二氧杂环丁烷作为化学发光源的印迹测定使用由至少一种式（1）单体组成的聚合物作为改进的膜。 膜减少背景信号，提高灵敏度和可靠性。

公开(公告)号：WO1998027105A1

公开(公告)日：1998-06-25

申请号：PCT/US1997022138

申请日：1997-12-11

Applicant: TROPIX, INC.

Inventor： TROPIX, INC. ,  BRONSTEIN, Irena ,  EDWARDS, Brooks ,  MARTIN, Christopher ,  VOYTA, John

IPC: C07J01/00

CPC classification number: C07H21/00 ,  C07D321/00 ,  C12Q1/6816 ,  C12Q1/686 ,  Y02P20/55 ,  C12Q2561/101 ,  C12Q2565/113

Abstract: Probes labeled with 1,2-dioxetane precursors can be employed in a variety of assays. The probes may be nucleic acid, peptide nucleic acid, proteins including enzyme, antibody or antigen, steroid, carbohydrate, drug or non-drug hapten. The probe is provided with a 1,2-dioxetane precursor bound thereto, generally either covalently, or a strong ligand bond. The dioxetane precursor moiety is converted to a bound 1,2-dioxetane by exposure to singlet oxygen. The dioxetane (labels) either spontaneously decompose, or are induced to decompose by an appropriate trigger to release light. The trigger may be a change in pH temperature, or an agent which removes a protective group. Assay formats in which these 1,2-dioxetane labeled probes and referents may be used to include hybridization assays, immuno assays, gel-based assays and Capillary Zone Electrophoresis.

Abstract translation: 用1,2-二氧杂环丁烷前体标记的探针可用于多种测定。 探针可以是核酸，肽核酸，蛋白质，包括酶，抗体或抗原，类固醇，碳水化合物，药物或非药物半抗原。 探针具有与其结合的1,2-二氧杂环丁烷前体，通常共价或强配体键。 通过暴露于单线态氧将二氧杂环丁烷前体部分转化为结合的1,2-二氧环乙烷。 二氧杂环丁烷（标记）自发分解，或通过适当的触发剂诱发分解以释放光。 触发可能是pH温度的变化，或者是除去保护基团的试剂。 这些1,2-二氧杂环丁烷标记的探针和指示物可用于包括杂交测定，免疫测定，基于凝胶的测定和毛细管区域电泳的测定形式。

公开(公告)号：WO1994026726A1

公开(公告)日：1994-11-24

申请号：PCT/US1994004555

申请日：1994-05-06

Applicant: TROPIX, INC.

Inventor： TROPIX, INC. ,  BRONSTEIN, Irena ,  EDWARDS, Brooks ,  SPARKS, Alison

IPC: C07D265/00

CPC classification number: G01N33/582 ,  A43B1/0036 ,  A43B1/0072 ,  A43B3/0031 ,  C07C43/1788 ,  C07C43/196 ,  C07C43/215 ,  C07C43/225 ,  C07C43/23 ,  C07C43/315 ,  C07C45/673 ,  C07C47/565 ,  C07C47/575 ,  C07C65/03 ,  C07C65/21 ,  C07C205/37 ,  C07C205/44 ,  C07C217/84 ,  C07C233/18 ,  C07C271/28 ,  C07C271/58 ,  C07C2603/74 ,  C07D321/00 ,  C07F9/12 ,  C07F9/4056 ,  C07F9/65512 ,  C07F9/65742 ,  C07G3/00 ,  C07H15/203 ,  C07H15/26 ,  C12Q1/34 ,  C12Q1/37 ,  C12Q1/42 ,  C12Q2334/00 ,  G01N21/76 ,  G01N33/581 ,  Y10S435/81 ,  Y10S435/975

Abstract: Novel 1,2-dioxetanes with improved chemiluminescent properties, such as signal intensity, S/N ratio, T1/2, etc. are provided by spiroadamantyl 1,2-dioxetanes, wherein the remaining carbon atom of the ring bears an alkoxy, aryloxy, or arylalkoxy substituent, and either a phenyl or naphthyl ring, this aromatic ring bearing, at the meta position on the phenyl group, or a non-conjugated position on the naphthyl ring, an OX moiety wherein X is an enzyme-clevable group, which when removed from the dioxetane, leaves the oxyanion which decomposes with chemiluminescence, the aryl ring further bearing an electron-active substituent Z. The nature and placement of the Z substituent, at a position not adjacent the point of attachment to the dioxetane ring, strongly influences the properties of the dioxetane. Assays, as well as kits for the performance of those assays, include the dioxetane, an enzyme capable of cleaving the X group, and in certain cases, membranes and chemiluminescent enhancement agents.

Abstract translation: 具有改善的化学发光性能的新型1,2-二氧杂环丁烷，如信号强度，S / N比，T1 / 2等，由金刚烷基1,2-二氧杂环丁烷提供，其中环的剩余碳原子为烷氧基，芳氧基 或芳基烷氧基取代基，苯基或萘基环，该芳环在苯基间位，或在萘环上的非结合位置，O X部分，其中X是可酶解的基团， 当从二氧杂环丁烷中除去时，离开化学发光分解的氧阴离子，芳基还具有电子活性取代基Z.Z-取代基在不与二氧杂环丁烷环的连接点相邻的位置处的性质和位置， 强烈地影响二氧杂环丁烷的性质。 测定以及用于进行这些测定的试剂盒包括二氧杂环丁烷，能够切割X基团的酶，以及在某些情况下，包括膜和化学发光增强剂。

公开(公告)号：WO1990000164A1

公开(公告)日：1990-01-11

申请号：PCT/US1989002793

申请日：1989-06-28

Applicant: TROPIX, INC.

Inventor： TROPIX, INC. ,  EDWARDS, Brooks ,  BRONSTEIN, Irena, Y. ,  LAIRD, Alison, A. ,  VOYTA, John, C.

IPC: C07C39/14

CPC classification number: C07C43/215 ,  C07C43/225 ,  C07C43/23 ,  C07C49/84 ,  C07C69/00 ,  C07C2603/00 ,  C07C2603/74 ,  C07D321/00 ,  C07F9/65512 ,  C12Q1/34 ,  G01N33/582 ,  Y10S435/968 ,  Y10S435/973

Abstract: Chemiluminescent 1,2-dioxetane compounds are disclosed in which the molecule is stabilized at the 3-position on the dioxetane ring against decomposition prior to the molecule's coming in contact with a labile group-removing substance and substituted at the 4-position on the dioxetane ring with a fused polycyclic ring-containing fluorophore moiety bearing a labile ring substituent whose point of attachment to the fused polycyclic ring, in relation to this ring's point(s) of attachment to the dioxetane ring, is such that the total number of ring atoms separating these points of attachment, including the ring atoms at the points of attachment, is an odd whole number. They are useful in detecting the presence or determining the concentration of chemical or biological substances in immunoassays, chemical assays and nucleic acid probe assays.

公开(公告)号：WO1997024460A1

公开(公告)日：1997-07-10

申请号：PCT/US1996020650

申请日：1996-12-23

Applicant: TROPIX, INC.

Inventor： TROPIX, INC. ,  BRONSTEIN, Irena, Y. ,  FORTIN, John, J. ,  MARTIN, Chris, S. ,  VOYTA, John, C.

IPC: C12Q01/68

CPC classification number: C12Q1/34 ,  C12Q1/6897

Abstract: A method of measuring the activity of at least two reporter gene products in an aliquot of a sample extract is disclosed. The activities of a first and second reporter enzyme are quantified by measuring the light signal produced by degradation of a first substrate by the first reporter enzyme and the light signal produced by the degradation of a second substrate by a second reporter enzyme. Both quantifications are sequentially performed on the same aliquot of sample extract.

Abstract translation: 公开了一种测量样品提取物等分试样中至少两种报告基因产物的活性的方法。 通过测量由第一报道酶降解第一底物产生的光信号和通过第二报道酶对第二底物的降解产生的光信号来量化第一和第二报道酶的活性。 在相同等分的样品提取物上依次进行两个定量。

公开(公告)号：WO1996024849A1

公开(公告)日：1996-08-15

申请号：PCT/US1996000035

申请日：1996-01-22

Applicant: TROPIX, INC.

Inventor： TROPIX, INC. ,  BRONSTEIN, Irena ,  EDWARDS, Brooks ,  MARTIN, Christopher ,  SPARKS, Alison ,  VOYTA, John, C.

IPC: G01N33/573

CPC classification number: C07K5/06095 ,  C07D321/00 ,  C07K5/0802 ,  C07K5/0817 ,  C07K5/1002 ,  C12Q1/37 ,  G01N33/582 ,  Y10S530/807

Abstract: 1,2-Dioxetane compounds bearing a proteolytic enzyme specific amino acid or peptide are provided, which amino acid or peptide can be removed by action of the corresponding protease. When the amino acid or peptide is removed, the 1,2-dioxetane decomposes with chemiluminescence, the generation of light providing a rapid, ultra-sensitive and convenient means for detecting the presence of the protease in the sample being inspected. The amount of light generated, or degree of chemiluminescence, can be correlated with the amount of protease present. Immunoassays, as well as DNA hybridization and DNA probe assays are provided.

Abstract translation: 提供了带有蛋白水解酶特异性氨基酸或肽的1,2-二氧杂环丁烷化合物，该氨基酸或肽可以通过相应的蛋白酶的作用除去。 当去除氨基酸或肽时，1,2-二氧杂环丁烷以化学发光分解，产生光，提供快速，超灵敏和方便的手段，用于检测待检样品中蛋白酶的存在。 产生的光量或化学发光度可以与存在的蛋白酶的量相关。 提供免疫测定，以及DNA杂交和DNA探针测定。