Non Classical Platinum Drugs: The Trans Isomers.people.unica.it/valeriamnurchi/files/2015/06/Natile-1.pdf · Non Classical Platinum Drugs: The Trans Isomers. Giovanni NATILE ... Mean

Non Classical Platinum Drugs: The Trans Isomers.

Giovanni NATILE

Chemistry Department, University of Bari via E. Orabona 4, 70125 Bari, Italy [email protected]

CISPLATIN

(CDDP)

PLATINUM COMPOUNDS IN CLINICAL USE

LIMITATIONS

Narrow spectrum of activity

Toxicity (particularly nephro- and neuro-

toxicity)

Resistance (intrinsic or acquired)

Development of new platinum compounds Wider

spectrum of

activity

Better

Pharmacology

New platinum

drugs

Cl

Cl N

N

Cl

Cl

Cl

Cl NH3 Pt

NH3

- Bone tissue affinity

- Inhibition of metalloproteinases

- Activation of trans geometry

- Different mode of DNA interaction

Different ligands for new antitumoral platinum compouds

Ketimines S

O

P

O

O

O (SMP)

C N H

OCH 3

CH 3

Iminoethers P

O

P

O

O

O

O

O H

H H

H

R R1

Bisphosphonate

C N H

NH2

CH 3

Amidines

C N H

CH 3

CH 3

H2N P

O

O

O (AMP)

P t

H 3 N C l

C l N

C H 3 H

O C H 3

C

t r a n s - Z

N C

O C H 3 H

C H 3 P t

C l

C l

N C

O H 3 C H

H 3 C

t r a n s - E E

P t

N H 3

C l N

C l

H 3

t r a n splatin

P t

N H 3

C l N

C H 3 H

O C H 3

C

C l

c i s - Z

P t

C l

N C

O C H 3 H

C H 3

C l

N C

O C H 3 H

C H 3

c i s - E E c i splatin

P t

N H 3

C l N

C l

H 3

Mean IC 50 ( m M) and Range (IC 50(least sens.) /IC 50(most sen.) ) in a panel of 6 human tumor cell lines

Complex IC 50 Range trans-EE 2.4 12 cis-EE 30 5 cis-DDP 4.6 25 trans-DDP 124 4

Antileukemic effect (P388 system) of platinum complexes

Complex in vitro in vivo

ID 50 ( m M) Dose % T/C

(mg/kg, qd 1-7) cis-DDP 2 0.6 211 cis-EE 7.7 4 150

- 8 toxic trans-EE 2.2 8 171

- 12 196

ca07

Control & dienPt

cisPt

Trans-EE

PRIMER EXTENSION ACTIVITY OF EXONUCLEASE-DEFICIENT KLENOW FRAGMENT OF DNA POLYMERASE I (KF-)

INTERACTION OF PLATINATED DNA WITH DOMAINS A AND B OF HMGB1

TERNARY DNA-PROTEIN COMPLEX FORMATION WITH: (A) THE SEQUENCE-SPECIFIC DNA-BINDING PROTEIN ECORI RESTRICTION ENDONUCLEASE,

(B) THE NON-SEQUENCE-SPECIFIC DNA-BINDING PROTEIN, LINKER HISTONE H1

PRIMER EXTENSION ACTIVITY OF: (A) EXONUCLEASE-DEFICIENT KLENOW FRAGMENT OF DNA POLYMERASE I (KF-)

(B) REVERSE TRANSCRIPTASE FROM HUMAN IMMUNODEFICIENCY VIRUS 1 (RT HIV-1)

A

B

EXCISION OF THE ADDUCTS OF PLATINUM COMPLEXES BY RODENT EXCINUCLEASE (THE PLATINATED 148 BP SUBSTRATE INCUBATED WITH CHO AA8 CFE FOR 40 MIN AT 30 °C)

- Trans-compounds retain their carrier ligands when

they are bound to MT

- The rate of Zn(II) release from MT by different Pt(II)

compounds is diverse and may affect free Zn(II)-

related pathways (MTF1, p53, reactive oxygen species)

Knipp, Karotki, Chesnov, Natile, Sadler, Brabec, Vasak, J. Med. Chem., 2007

Cytochrome c (cyt c)

Fe(III)

heme

Met65

His33 His26

His18

Met80

-Small heme protein found in the

mitochondrion

-It transfers electrons between

Complexes III and IV of the

respiratory chain

-It is a critical protein in the apoptotic

pathway

H3N

Pt

Cl

Cl

H3N

+ Cytc+ 2L

- NH3

- 2Cl

L

Pt

Cytc

L

H3N - L

L

Pt

H3N

Cytc

Casini, Gabbiani, Mastrobuoni, Pellicani, Intini, Arnesano, Natile, Moneti,

Francese, Messori, Biochemistry, 2007

Cl

Pt

Cl

ImE

ImE

+ Cytc

- Cl

Cl

Pt

Cytc

ImE

ImE- Cl

Pt

ImE

CytcImE

Thioether can Serve as Intermediates for DNA Platination

by trans Geometry Antitumor Drug

C. Li, Z. Li, E. Sletten, F. Arnesano, M. Losacco, G. Natile, Y. Liu, Angew. Chem. 2009, 48, 8497

Pt

ClE

Cl E

+ Met

high NaCl

low NaCl

Pt

ClE

E

Pt

E

EMet-S

S-Met

very fast

fast

Pt

E

EMet-S

GMP

N7-GMPMet-S

GMP

Effect of thioethers on DNA platination by trans-Platinum Complexes

C. Li, R. Huang, Y. Ding, E. Sletten, F. Arnesano, M. Losacco, G. Natile, Y. Liu, Inorg. Chem. 2011, 50, 8168

0

100

200

300

IC50

(µM

)

cisplatin ampyplatincDPCP transplatin

A549RA549

MCF-7HeLa

ampyplatin

Pt

NH3

N

ClHMG

protein

PtCl

NH3

N

Cl

D. Xu, Y. Min, Q. Cheng, H. Shi, K. Wei, F. Arnesano, G. Natile, Y. Liu, J. Inorg. Biochem., in press

Interesting chemistry of imine complexes: synthesis of cis-[PtCl2{Z-HN=C(But)NH2}2] and cis-[PtCl2NH3{Z-HN=C(But)NH2}]

Pt

Cl

L

Cl

N

C

But

Pt

Cl

L

Cl

N

H

NH2But

THF, 55 °C

NH3 aq (1:5)

L = NH3

N C ButL =

L = NH3

L= HN C

NH2

But

Synthesis of trans-[PtCl2{Z-HN=C(But)NH2}2] and trans-[PtCl2NH3{Z-HN=C(But)NH2}]

Pt

Cl

Cl

L

N

C

But

Cl2 / CCl4

CHCl3, 25 ° C

Pt

Cl

Cl

Cl

Cl

L

N

C

But

CH2Cl2, 25 ° C

NH3 aq (1:5)

Pt

Cl

Cl

Cl

Cl

L

NH

NH2But

ascorbic acid (1:3)

H2O, 25 ° C

Pt

Cl

Cl

L

N

NH2But

H

L = NH3

N C ButL =

L = NH3

L= HN C

NH2

But

L = NH3

L= HN C

NH2

But

L = NH3

N C ButL =

X-Ray crystal structure of cis-[PtCl2NH3{Z-HN=C(But)NH2}]

2.53 Å 3.4 Å 2.65 Å

In vitro antitumor activity

S.D. = standard deviation IC50 values were calculated by probit analysis (P < 0.05, X2 test). Cells (3-8·104·ml-1) were treated for 48 hours with increasing concentrations of tested compounds dissolved in PEG400. Cytotoxicity was assessed by MTT test.

The prepared complexes were tested in vitro against 8 human cancer cell lines. The compound with trans configuration trans-1d is the most active. Both trans-1d or cis-1d are more active than cisplatin, while complexes trans-2d and cis-1d have cytotoxic activity comparable (or slightly lower) than that of cisplatin.

Compound IC50 (µM) ± S.D.

MCF-7 A375 A549 HCT-15 LoVo

cis-[PtCl2{Z-HN=C(NH2)But}2] 24.97±1.05 39.25±1.05 36.54±2.37 35.14±1.43 24.34±2.00

trans-[PtCl2{Z-HN=C(NH2)But}2] 23.86±3.63 27.12±2.74 24.57±2.01 36.33±1.93 27.05±1.64

cis-[PtCl2(NH3){Z-HN=C(NH2)But}] 18.25±1.24 17.26±2.41 15.32±2.61 16.98±2.21 15.55±1.38

trans-[PtCl2(NH3){Z-HN=C(NH2)But}] 7.15±1.27 7.54±1.87 6.62±1.45 11.51±1.75 7.12±2.12

[PtIII

2Cl2I2{HN=C(But)NH}2(NH3)2] (2SM189A) 27.67±3.14 23.46±2.38 19.64±3.26 18.52±3.16 21.16±3.04

trans-[PtCl4(NH3){Z-HN=C(But)NH2}]

(2SM106B) 46.64±3.15 41.12 ±3.75 56. 16±4.14 51.50±3.26 49.19±4.06

Cisplatin 25.18±2.58 18.28±1.05 25.45±1.09 20.31±2.13 15.9±1.50

cis-[PtCl2{Z-HN=C(But)NH

2}

2], cis-2d

trans-[PtCl2{Z-HN=C(But)NH

2}

2], trans-2d

trans-[PtCl2(NH

3){Z-HN=C(But)NH

2}

2], trans-1d

cis-[PtCl2(NH

3){Z-HN=C(But)NH

2}

2], cis-1d

trans-[PtCl4(NH

3){Z-HN=C(But)NH

2}

2], trans-1c

3e







[PtIII



(2SM106B) 46.64±3.15 41.12 ±3.75 56. 16±4.14 51.50±3.26 49.19±4.06

Cisplatin 25.18±2.58 18.28±1.05 25.45±1.09 20.31±2.13 15.9±1.50


2}

2], cis-2d


2}

2], trans-2d

trans-[PtCl2(NH

3){Z-HN=C(But)NH

2}

2], trans-1d

cis-[PtCl2(NH

3){Z-HN=C(But)NH

2}

2], cis-1d

trans-[PtCl4(NH

3){Z-HN=C(But)NH

2}

2], trans-1c







[PtIII



(2SM106B) 46.64±3.15 41.12 ±3.75 56. 16±4.14 51.50±3.26 49.19±4.06

Cisplatin 25.18±2.58 18.28±1.05 25.45±1.09 20.31±2.13 15.9±1.50


2}

2], cis-2d


2}

2], trans-2d

trans-[PtCl2(NH

3){Z-HN=C(But)NH

2}

2], trans-1d

cis-[PtCl2(NH

3){Z-HN=C(But)NH

2}

2], cis-1d

trans-[PtCl4(NH

3){Z-HN=C(But)NH

2}

2], trans-1c

3e

cis-[PtCl2(NH3){Z-HN=C(But)NH2}], cis-1d

trans-[PtCl2(NH3){Z-HN=C(But)NH2}], trans-1d

trans-[PtCl4(NH3){Z-HN=C(But)NH2}], trans-1c

cis-[PtCl2{Z-HN=C(But)NH2}2], cis-2d

trans-[PtCl2{Z-HN=C(But)NH2}2], trans-2d

Cross-resistance profiles in ovarian cancer cells

S.D. = standard deviation IC50 values were calculated by probit analysis (P < 0.05, X2 test). Cells (3-8·104·ml-1) were treated for 48 hrs with increasing concentrations of tested compounds dissolved in PEG400. Cytotoxicity was assessed by MTT test. R.F.= IC50 resistant/ IC50 sensitive cells.

The RF value was about 7 times lower than that calculated for cisplatin. These results clearly indicated that these complexes possesses a different cross-resistance profile than that of cisplatin.

Human ovarian adenocarcinoma cells

Compound 2008

IC50 (µM) ± S.D.

C13

IC50 (µM) ± S.D.

R.F.

cis-[PtCl2{Z-HN=C(NH2)But}2] 22.34±1.97 23.54±2.14 1.1

trans-[PtCl2{Z-HN=C(NH2)But}2] 25.11±2.32 31.85±1.80 1.3

cis-[PtCl2(NH3){Z-HN=C(NH2)But}] 12.56±2.76 20.57±1.55 1.0

trans-[PtCl2(NH3){Z-HN=C(NH2)But}] 6.11±2.09 7.61±1.25 1.2

[PtIII

2Cl2I2{HN=C(But)NH}2(NH3)2]. 23.14±2.41 26.63±2.95 1.1

Trans-[PtCl4(NH3){Z-HN=C(But)NH2}] 46.52±3.18 63.34±4.25 1.4

Cisplatin 12.69±1.72 89.18±1.51 7.03


2}

2], cis-2d


2}

2], trans-2d

trans-[PtCl2(NH

3){Z-HN=C(But)NH

2}

2], trans-1d

cis-[PtCl2(NH

3){Z-HN=C(But)NH

2}

2], cis-1d

trans-[PtCl4(NH

3){Z-HN=C(But)NH

2}

2], trans-1c

3e


Compound 2008

IC50 (µM) ± S.D.

C13

IC50 (µM) ± S.D.

R.F.





[PtIII

2Cl2I2{HN=C(But)NH}2(NH3)2]. 23.14±2.41 26.63±2.95 1.1


Cisplatin 12.69±1.72 89.18±1.51 7.03


2}

2], cis-2d


2}

2], trans-2d

trans-[PtCl2(NH

3){Z-HN=C(But)NH

2}

2], trans-1d

cis-[PtCl2(NH

3){Z-HN=C(But)NH

2}

2], cis-1d

trans-[PtCl4(NH

3){Z-HN=C(But)NH

2}

2], trans-1c

3e


Compound 2008

IC50 (µM) ± S.D.

C13

IC50 (µM) ± S.D.

R.F.





[PtIII

2Cl2I2{HN=C(But)NH}2(NH3)2]. 23.14±2.41 26.63±2.95 1.1


Cisplatin 12.69±1.72 89.18±1.51 7.03


2}

2], cis-2d


2}

2], trans-2d

trans-[PtCl2(NH

3){Z-HN=C(But)NH

2}

2], trans-1d

cis-[PtCl2(NH

3){Z-HN=C(But)NH

2}

2], cis-1d

trans-[PtCl4(NH

3){Z-HN=C(But)NH

2}

2], trans-1c

3e


Compound 2008

IC50 (µM) ± S.D.

C13

IC50 (µM) ± S.D.

R.F.





[PtIII

2Cl2I2{HN=C(But)NH}2(NH3)2]. 23.14±2.41 26.63±2.95 1.1


Cisplatin 12.69±1.72 89.18±1.51 7.03


2}

2], cis-2d


2}

2], trans-2d

trans-[PtCl2(NH

3){Z-HN=C(But)NH

2}

2], trans-1d

cis-[PtCl2(NH

3){Z-HN=C(But)NH

2}

2], cis-1d

trans-[PtCl4(NH

3){Z-HN=C(But)NH

2}

2], trans-1c

3e

cis-[PtCl2(NH3){Z-HN=C(But)NH2}], cis-1d

trans-[PtCl2(NH3){Z-HN=C(But)NH2}], trans-1d

trans-[PtCl4(NH3){Z-HN=C(But)NH2}], trans-1c

cis-[PtCl2{Z-HN=C(But)NH2}2], cis-2d

trans-[PtCl2{Z-HN=C(But)NH2}2], trans-2d

Cisplatin

Documents

Non Classical Platinum Drugs: The Trans Isomers.people.unica.it/valeriamnurchi/files/2015/06/Natile-1.pdf · Non Classical Platinum Drugs: The Trans Isomers. Giovanni NATILE ... Mean