Chromogenic enzyme substrates
    6.
    发明授权
    Chromogenic enzyme substrates 失效
    显色酶底物

    公开(公告)号:US4252715A

    公开(公告)日:1981-02-24

    申请号:US54075

    申请日:1979-07-02

    摘要: New chromogenic substrates suitable for serine proteases, specially suitable for diagnostic determination of factor Xa, which are represented by the following formula:R.sub.1 -A.sub.1 -A.sub.2 -Gly-Arg-NH-R.sub.2or its salts, where R.sub.1 is selected from the group of hydrogen, alkanoyl having from 1 to 12 carbon atoms, cyclohexylcarbonyl, benzoyl, benzoyl substituted with one or two halogen atoms, methylamine or phenyl groups, benzene sulphonyl and toluenesulphonyl; R.sub.2 is a chromophoric group; A.sub.1 is selected from the group of a single bond, and the amino acids selected from the group of Gly, Ala, Val, Leu, Ileu, Pro, Met, Phe and Tyr; and A.sub.2 is selected from the group of the amino acids Glu, Gln, Asp, and Asn.

    摘要翻译: 适用于丝氨酸蛋白酶的新显色底物,特别适用于由下式表示的因子Xa的诊断测定:R1-A1-A2-Gly-Arg-NH-R2或其盐,其中R1选自 氢,具有1至12个碳原子的烷酰基,环己基羰基,苯甲酰基,被一个或两个卤素原子取代的苯甲酰基,甲胺或苯基,苯磺酰基和甲苯磺酰基; R2是发色团; A1选自单键,氨基酸选自Gly,Ala,Val,Leu,Ileu,Pro,Met,Phe和Tyr; 并且A2选自氨基酸Glu,Gln,Asp和Asn的组。

    Determination of serine proteases
    7.
    发明授权
    Determination of serine proteases 失效
    丝氨酸蛋白酶的测定

    公开(公告)号:US4276375A

    公开(公告)日:1981-06-30

    申请号:US86970

    申请日:1979-10-22

    摘要: Serine proteases in a substance such as blood are determined by contacting the substance with a novel chromogenic or fluorgenic substrate for serine proteinases and spectrophotometrically measuring the quantity of a chromophore or fluorescent compound released from the substrate by serine proteases in the substance. The novel substrate has the amino acid sequence-ILe-A-Gly-Arg-wherein A is Asp or Glu substituted in the carboxylic group by esterification or amidation. The novel substrate increases sensitivity and accuracy of the determination.

    摘要翻译: 通过使该物质与丝氨酸蛋白酶的新的显色或荧光底物接触并通过分光光度法测量从物质中丝氨酸蛋白酶从底物释放的发色团或荧光化合物的量来测定血液中的丝氨酸蛋白酶。 该新型底物具有氨基酸序列-Ie-A-Gly-Arg-,其中A是通过酯化或酰胺化在羧基中被取代的Asp或Glu。 新颖的底物提高了测定的灵敏度和准确性。

    Method for determining a proteolytic enzyme
    8.
    发明授权
    Method for determining a proteolytic enzyme 失效
    测定蛋白水解酶的方法

    公开(公告)号:US4162941A

    公开(公告)日:1979-07-31

    申请号:US774350

    申请日:1977-03-04

    IPC分类号: C12Q1/56 G01N31/14

    摘要: The proteolytic enzyme designated as factor Xa is diagnostically determined by contacting the enzyme with a chromogenic substrate represented by the formulaR.sub.1 -A.sub.1 -A.sub.2 -Gly-Arg-NH-R.sub.2or its salts, where R.sub.1 is hydrogen or alkanoyl having from 1 to 12 carbon atoms or cyclohexylcarbonyl or benzoyl or benzoyl substituted with one or two halogen atoms, methylamine or phenyl groups or benzene sulphonyl or toluenesulphonyl, R.sub.2 is nitrophenyl or naphthyl or nitronaphthyl or methoxynaphthyl or quinolyl or nitroquinolyl, A.sub.1 is a single bond or one of the amino acids Gly, Ala, Val, Leu, Ileu, Pro, Met, Phe or Tyr, and A.sub.2 is one of the amino acids Glu, Gln, Asp, or Asn and spectrophotometrically measuring the degree of enzymatic hydrolysis.

    摘要翻译: 通过使酶与由式R1-A1-A2-Gly-Arg-NH-R2或其盐表示的显色底物接触而诊断性地确定为因子Xa的蛋白水解酶,其中R 1为氢或具有1至12个的烷酰基 碳原子或环己基羰基或被一个或两个卤素原子取代的苯甲酰基或苯甲酰基,甲胺或苯基或苯磺酰基或甲苯磺酰基,R2是硝基苯基或萘基或硝基萘基或甲氧基萘基或喹啉基或硝基奎啉基,A1是单键或氨基 酸,Gly,Ala,Val,Leu,Ileu,Pro,Met,Phe或Tyr,A2是氨基酸Glu,Gln,Asp或Asn之一,并通过分光光度法测量酶水解度。

    Peptide substrates for determination of protease activity

    公开(公告)号:US4748116A

    公开(公告)日:1988-05-31

    申请号:US53569

    申请日:1987-05-21

    摘要: Peptide sequences consisting of 2-4 amino acids with high affinity and comparatively high specificity to a number of various, physiologically important proteases are known to have been synthetized before. Such sequences with an added C-terminal marker have been widely used as substrates for the quantitative determination of the kind of proteases mentioned above. The method is based on the fact that the marker is split off under influence of the enzyme and that the liberated marker possesses an easily measurable, for instance, optic property which differs from that of the original substrate. The type of markers used until today have mainly been chromophores or fluorophores which can be quantified by photometry or fluorometry.The present invention relates to a new type of markers coupled to known peptide sequences. These markers are luminol or isoluminol which in their free state can be brought to luminate intensely, but lose considerably in luminescence when they are amide-bound to a peptide sequence.The peptide derivatives consist of acyl derivates of luminol (5-amino-2,3-dihydro-1,4-phthalazinedione) or isoluminol (6-amino-2,3-dihydro-1,4-phthalazinedione) where the acyl residue consists of an amide-bound amino acid or amino acid sequence with 2-4 amino acid residues and where the .alpha.-amino group is either free of acylated.The greatest advantages of the luminogenic substrates according to the invention are that theyare considerably more sensitive than the previously use chromogenic or fluorogenic substrates and can be used for measuring even extremely low protease concentration;permit measurement of minute final volumes in standard luminometers which also are technically less complicated and, therefore, cheaper than both spectrophotometers and fluorometers; as a result, costs for both analysis reagents and devices can be cut down;permit linear response measurements within a much wider concentration range than is the case with the usual chromogenic and fluorogenic methods;generate markers less sensitive to deactivating interference as, for instance, chemical quenching; this distinguishes them from, for instance, fluorogenic substrates.