Pt-containing compound, process for its preparation, and application of such compounds

• US 5,580,990 A
• Filed: 10/29/1993
• Issued: 12/03/1996
• Est. Priority Date: 07/19/1990
• Status: Expired due to Term

• Current Assignee: Rijksuniversiteit te Leiden, Stichting Klinische Research Academisch Medisch Centrum
• Original Assignee: Rijksuniversiteit te Leiden, Stichting Klinische Research Academisch Medisch Centrum
• Inventors: Reedijk, Jan, van den Berg, Franciscus M., Lempers, Edwin L. M., Bloemink, Marieke J.
• Primary Examiner(s): Shah, Mukund J.
• Assistant Examiner(s): WONG, KING
• Application Number:
  US07/975,586
• Time in Patent Office: 1,131 Days
• Field of Search: 435/5, 435/6, 556/137, 424/9.36
• US Class Current: 549/212
• CPC Class Codes: C07F 15/0093 without a metal-carbon linkage
• CPC Class Codes: C07J 51/00 Normal steroids with unmodified cyclopenta(a)hydrophenanthrene skeleton not provided for in groups C07J1/00 - C07J43/00
• CPC Class Codes: C09B 11/04 derived from triarylmethanes , i.e. central C-atom is substituted by amino, cyano, alkyl
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 2563/137 Metal/ion, e.g. metal label

Abstract

Pt-containing compound with the formula {Pt^II (w)(x)(y)(z)} or {Pt^IV (u)(v)(w)(x)(y)(z)}, wherein u, v, w, x, y and z represent whether or not the same whether or not interconnected ligands, of which at least one is a leaving ligand and at least one of the remaining ligands represents a detectable marker group. According to the invention (CH₃)₂ SO, Cl or H₂ O appears to be suitable as a leaving ligand while as detectable marker group a fluorescent group merits preference and is the ligand fluorescein or tetramethyl rhodamine. A suitable Pt-containing compounds is {Pt(ethylenediamine)(Me₂ SO)(fluorescein-NH(CS)-NHCH₃)} (PtF). Further, the invention comprises a process for the preparation of the Pt-containing compounds, where the Pt-containing compounds are prepared in a manner known per se for analogous compounds.

Claims

1. Pt-containing compound for use in detectably labeling a nucleic acid molecule, with the formula {Pt^II (w)(x)(y)(z)} or {Pt^IV (u)(v)(w)(x)(y)(z)}, with the structural formula 1 or 2:

##STR4## in which u, v, w, x, y, and z represent whether or not the same whether or not covalently interconnected ligands, of which no more than one of the ligands is a leaving ligand selected from the group consisting of halide, sulfate, nitrate, phosphate, carbonate, ethylnitrate, phosphonates, oxalates, citrates, H₂ O ROH, RO^-, in which R is an organic residual group; and

substituted sulfoxides R¹ R² SO, in which R¹ and R² whether or not equal to each other are organic residual groups, at least one of the ligands is a detectable marker group, and the remaining ligands are one or more ethylenediamine moieties.

2. Compound according to claim 1, wherein the leaving ligand is (CH₃)₂ SO, Cl, or H₂ O.

3. Compound according to claim 1, wherein the detectable marker group is a fluorescent group.

4. Compound according to claim 3, wherein the fluorescent group is fluorescein or tetramethyl rhodamine, or either compound optionally modified to include a Pt-binding linkage structure containing a double-bonded S atom, a --SR group, a NR³ R⁴ group or a nitrogen-containing ring, wherein R is an organic residual group and R³ and R⁴ are independently hydrogen atoms or organic residual groups.

5. The Pt-containing compound according to claim 1, wherein the detectable marker group is fluorescein, rhodamine, 7-amino-4-methylcoumarin-3-acetic acid, biotin or other hapten, digoxygenin, a radioactive moiety, or an immunohistochemically detectable moiety, and wherein said detectable marker group is optionally modified to include a Pt-binding linkage structure containing a double-bonded S atom, a --SR group, a NR³ R⁴ group or a nitrogen-containing ring, wherein R is an organic residual group and R³ and R⁴ are independently hydrogen atoms or organic residual groups.

6. The Pt-containing compound according to claim 1, wherein the detectable marker group is biotin optionally modified to include a Pt-binding linkage structure containing a double-bonded S atom, a --SR group, a NR³ R⁴ group or a nitrogen-containing ring, wherein R is an organic residual group and R³ and R⁴ are independently hydrogen atoms or organic residual groups.

7. The Pt-containing compound according to claim 1, having the structure:

##STR5## wherein w is said leaving groups, x is biotin optionally modified to include a Pt-binding linkage structure containing a double-bonded S atom, a --SR group, a NR³ R⁴ group or a nitrogen-containing ring, wherein R is an organic residual group and R³ and R⁴ are independently hydrogen atoms or organic residual groups, and y and z together are ethylenediamine.

8. {Pt(ethylenediamine)Cl(fluorescein-NH(CS)NHCH₃)}.

9. {Pt(ethylenediamine)(Me₂ SO)(fluorescein-NH(CS)-NHCH₃)}.

10. A process for preparing the compound of claim 9, comprising:

(a) reacting fluorescein-N═

C═

S with CH₃ NH₂ in water to produce fluorescein-NH(CS)NHCH₃ ;

(b) acidifying said fluorescein-NH(CS)NHCH₃ to a pH of about 2-3 to precipitate said fluorescein-NH(CS)NHCH₃ ;

(c) resuspending said precipitate in water;

(d) adjusting the pH of the suspension of fluorescein-NH(CS)NHCH₃ to about 10-11 to obtain a bright yellow solution;

(e) reacting the basified fluorescein-NH(CS)NHCH₃ with Pt(ethylenediamine)(Me₂ SO)Cl in water while stirring at room temperature in the dark;

(f) precipitating the non-reacted fluorescein-NH(CS)NHCH₃ by adjusting the pH of the reaction mixture to about 2-3; and

(g) filtering and freeze-drying the supernatant from step (f), yielding

Pt(ethylenediamine)(Me₂ SO)(fluorescein-NH(CS)-NHCH₃).

11. A method for the diagnosis of viral, bacterial, or parasitic infections, detection of genetic deviation, or detection of gene expression, comprising detecting a detectably labeled nucleic acid specific for a viral, bacterial, or parasitic infection, genetic deviation, or gene expression, wherein said detectably labeled nucleic acid is labeled with a Pt-containing compound with the formula {Pt^II (w)(x)(y)(z)} or {Pt^IV (u)(v)(w)(x)(y)(z)}, with the structural formula 1 or 2:

##STR6## in which u, v, w, x, y and z represent whether or not the same whether or not covalently interconnected ligands, of which no more than one of the ligands is said nucleic acid molecule, at least one of the ligands is a detectable marker group, and the remaining ligands are one or more ethylenediamine moieties.

12. The method according to claim 11, wherein the detectable marker group is a fluorescent group.

13. The method according to claim 12, wherein the fluorescent group is fluorescein or tetramethyl rhodamine or either compound optionally modified to include a Pt-binding linkage structure containing a double-bonded S atom, a --SR group, a NR³ R⁴ group or a nitrogen-containing ring, wherein R is an organic residual group and R³ and R⁴ are independently hydrogen atoms or organic residual groups.

14. The method according to claim 11, wherein the detectable marker group is fluorescein, rhodamine, 7-amino-3-methylcoumarin-3-acetic acid, biotin or other hapten, digoxygenin, a radioactive moiety, or an immunohistochemically detectable moiety, and wherein said detectable marker group is optionally modified to include a Pt-binding linkage structure containing a double-bonded S atom, a --SR group, a NR³ R⁴ group or a nitrogen-containing ring, wherein R is an organic residual group and R³ and R⁴ are independently hydrogen atoms or organic residual groups.

15. The method according to claim 11, wherein the detectable marker group is biotin optionally modified to include a Pt-binding linkage structure containing a double-bonded S atom, a --SR group, a NR³ R⁴ group or a nitrogen-containing ring, wherein R is an organic residual group and R³ and R⁴ are independently hydrogen atoms or organic residual groups.

16. The method according to claim 11, wherein said Pt-containing compound has the structure:

##STR7## wherein w is said nucleic acid molecule, x is biotin optionally modified to include a Pt-binding linkage structure containing a double-bonded S atom, a --SR group, a NR³ R⁴ group or a nitrogen-containing ring, wherein R is an organic residual group and R³ and R⁴ are independently hydrogen atoms or organic residual groups, and y and z together are ethylenediamine.