Multi-modal bioprobe for bladder cancer imaging and photodynamic therapy

Patent metadata

Field	Value
Publication number	US-9840522-B2
Application number	US-201615352561-A
Country	US
Kind code	B2
Filing date	Nov 15, 2016
Priority date	Nov 9, 2014
Publication date	Dec 12, 2017
Grant date	Dec 12, 2017

Abstract

The present invention relates to a new generation of PDT agents based on porphyrin-lanthanide complexes with specific functional groups which can specifically localize on particular tumors, and their PDT processes can be monitored via NIR emission from erbium. In particular, the present invention provides a multi-modal lanthanide-porphyrin PDT agent (Er—R 3 ) that are capable of killing the bladder tumor cells selectivity via 1 O 2 from porphyrin moiety and affording the fluorescence imaging simultaneously upon Er—R 3 binding with the integrin α v β 3 isoform in bladder cancer cells.

First claim

What we claim is:

1. A composition for photodynamic therapy and imaging of cancer cells comprising an Erbium porphyrin based complex, Ytterbium porphyrin based complex, or Gadolinium porphyrin based complex represented by the molecular formula:

wherein Ln is Er, Yb, or Gd; and
R n is a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 1; SEQ ID NO: 2; and SEQ ID NO: 3; or
a water-soluble porphyrin-based Gadolinium complex represented by a molecular formula selected from the group consisting of Gd 1 , Gd 2 , Gd 3 , Gd 4 , and Gd 5 :

or
a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 1; SEQ ID NO: 2; SEQ ID NO: 3; SEQ ID NO: 4; and SEQ ID NO: 5 or a pharmaceutically acceptable salt thereof.

2. The composition according to claim 1 wherein the Erbium porphyrin based complexes are conjugated with integrin α v β 3 isoform-specific peptides.

3. The composition according to claim 1 , wherein the composition comprises the Erbium porphyrin based complex and R n is SEQ ID NO: 4.

4. The composition according to claim 1 , wherein the composition comprises the Erbium porphyrin based complex and R n is SEQ ID NO: 5.

5. The composition according to claim 1 , wherein the composition comprises the Erbium porphyrin based complex and R n is SEQ ID NO: 3.

6. The composition according to claim 1 , wherein the composition comprises the Erbium porphyrin based complex represented by the molecular formula of:

wherein Ln is Er and R n is a polypeptide having an amino acid sequence of SEQ ID NO: 3.

7. The composition according to claim 1 wherein the cancer cells comprising of bladder cancer cells, cervical cancer cells and lung cancer.

8. A method of photodynamic therapy and imaging of cancer cells comprising administering to a subject in need thereof the composition according to claim 1 and irradiating the cancer cells in the subject in need thereof with a radiation source.

9. The method according to claim 8 wherein the administration of said composition is performed intravenously or by injection to site of said cancer cells.

10. The method according to claim 8 , wherein said radiation source is a light source with a wavelength in the Q band of porphyrin.

11. The method according to claim 8 , wherein said radiation source is a light source with a wavelength beyond 550 nm.

12. The method according to claim 8 , wherein said radiation source is a light source with a wavelength is at 860 nm.

13. The method according to claim 8 wherein the imaging is performed using fluorescent imaging.

14. The method according to claim 8 wherein the imaging is performed using NIR imaging.

15. The method according to claim 8 wherein the imaging is performed using MRI imaging.

16. The method according to claim 8 wherein the composition comprises the Gadolinium porphyrin based complex and Ln is Gd; or the composition comprises a compound selected from the group consisting of Gd 1 , Gd 2 , Gd 3 , Gd 4 , and Gd 5 .

17. A method of synthesizing the composition according to claim 1 wherein Ln=Er or Ln=Yb comprising steps according to the following scheme:

wherein
said compound Por(THP-TMS) is synthesized via steps comprising:
dissolving Pyrrole, pentafluorobenzaldehyde and 4-[2-(trimethylsilyl)ethynyl]benzaldehyde 6 in CH 2 Cl 2 under an argon atmosphere to produce a first solution;
leaving the first solution for at least 10 minutes;
adding BF 3 .O(Et) 2 to the first solution;
stirring the first solution for at least 1 hour at room temperature;
adding DDQ (2,3-Dichloro-5,6-dicyano-1,4-benzoquinone) to the first solution;
stirring the first solution for at least another 1 hour at room temperature;
removing the solvent from the first solution in vacuo to produce a first mixture;
passing the first mixture through a silica column (hexanes-CH 2 Cl 2 ) concentrated under reduced pressure to produce 5,10,15-Tris(pentafluorophenyl)-20-[4-{2-(trimethylsilyl)ethynyl}phenylporphyrin] or Por(THP-TMS);
said compound Ln-1 is synthesized via steps comprising:
dissolving Ln[N(SiMe 3 )2] 3 .x[LiCl(THF) 3 ]: HN(SiMe 3 ) 2 in THF at about 0 degrees Celcius to produce a second solution;
adding n-BuLi slowly over at least 30-minutes period to the second solution;
stirring the second solution for at least 12 hours;
transferring the second solution to a Schlenk flask with LnCl 3 suspended in THF to produce a second mixture;
stirring the second mixture for at least 24 hours until all of the solid LnCl 3 disappeared to produce Ln[N(SiMe 3 )2] 3 .x[Li(THF) 3 Cl] (x=3˜5) wherein Ln=Er or Ln=Yb;
said compound Yb-1 is further synthesized via steps comprising:
transferring Yb[N(SiMe 3 )2] 3 .x[Li(THF) 3 Cl] (x=3˜5) to a Schlenk flask;
removing the solvent from Yb[N(SiMe 3 )2] 3 .x[Li(THF) 3 Cl] (x=3˜5) under vacuum to produce a first residue;
adding CH 2 Cl 2 to the first residue for the precipitation of LiCl to produce a third mixture;
centrifuging the third mixture until a clear layer is produced;
transferring the clear layer to another Schlenk flask with dry Por(THP-TMS) free base dissolved in toluene to produce a third solution;
refluxing the third solution until most of the free base coordinated with the metal ion;
adding dry NaLOMe [LOMe-((cyclopentadienyl)tris(dimethylphosphito)-cobaltate or an anionic tripodalligand) to the third solution to produce a fourth mixture;
stirring the fourth mixture for at least another 12 hours;
cooling down the fourth mixture to room temperature;
removing the solvent from the fourth mixture in vacuum to produce a second residue;
dissolving the second residue in CHCl 3 ;
filtering and chromatographing the dissolved second residue on silica gel using CHCl 3 /petroleum ether as eluent;
further dissolving the output from chromatography in CH 2 Cl 2 ; and filtering the solution to produce compound Yb-1;
Said compound Er-1 is further synthesized via steps comprising:
the same steps as for Yb-1, replacing Yb[N(SiMe 3 )2] 3 .x[Li(THF) 3 Cl] (x=3˜5) with Er[N(SiMe 3 )2] 3 .x[Li(THF) 3 Cl] (x=3˜5);
said compound Ln-2 wherein Ln=Yr is synthesized via steps comprising:
adding TBAF to a solution of Yb-1 in CH 2 C12 to produce a fifth solution;
stirring the fifth solution for at least 30 minutes;
monitoring the progress of the reaction of the fifth solution by TLC;
after completion of the reaction, passing the fifth solution through a short of silica gel column;
removing the solvent from the fifth solution to produce Yr-2;
said compound Er-2 is further synthesized via

Assignees

• Univ Hong Kong Baptist Univ

Inventors

• Wong Ka Leung
• Wong Wai Kwok

Classifications

• A61N5/062 Primary

  Photodynamic therapy, i.e. excitation of an agent · CPC title

• C07F5/003 Primary

  without C-Metal linkages · CPC title

• A61K41/0071

  PDT with porphyrins having exactly 20 ring atoms, i.e. based on the non-expanded tetrapyrrolic ring system, e.g. bacteriochlorin, chlorin-e6, or phthalocyanines · CPC title

• C07F15/06

  Cobalt compounds · CPC title

• A61K41/008

  Two-Photon or Multi-Photon PDT, e.g. with upconverting dyes or photosensitisers · CPC title