steroidal platinum complexes

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

STEROIDAL PLATINUM COMPLEXES

DEPARTMENT OF MEDICINAL STEROIDS

INSTITUTE OF ORGANIC CHEMISTRY AND BIOCHEMISTRY

ACADEMY OF SCIENCES OF THE CZECH REPUBLIC

MIROSLAV KVASNICA

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

WHY PLATINUM ???

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

1. Diversity of possible complexes

PtH3N NH3

Cl Cl

cisplatin

PtO NH3

O NH3

O

Ocarboplatin

Pt

NH2

H2NO

O

O

O

oxaliplatin

PtHO OH

Cl Cl

NH2

NH2

iproplatin

PtH3N Cl

NH2

Cl

OH

OH

satraplatin

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

2. Using of platinum complexes

treatment of solid malignacies

(small cell lung cancer, ovarian and testicular cancer, epidermoid carcinomas of the head and neck, cancer of breast, uterus, and cervix)

Exhibits a wide spectrum of antitumour activity against drug-resistant as well as drug sensitive tumours

Shows activity against slow-growing tumour as well as rapidly-growing tumours

Shows no strain or species specificity

Exhibits activity against viral-induced and chemical-induced tumours

Affects both solid and disseminated tumours

Advantages of cisplatin

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

3. Mechanism of actioninside the cell: PtII(NH3)2Cl2 + H2O → [PtII(NH3)2Cl(H2O)]+ + Cl-

[PtII(NH3)2Cl(H2O)]+ + H2O → [PtII(NH3)2(H2O)2]2+

Pil, P., Lippard, S. J. In Encyclopedia of Cancer, J. R. Bertino, Ed. Academic Press: San Diego, CA, 1997, Vol. 1, pp. 392-410.

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

4. Drawbacks of cisplatin

toxicity

(nephrotoxicity, neurotoxicity, nausea and vomitting, ototoxicity, alopecia,

electrolyte disturbance, dermatitis)

drug resistance

* limit the formation of lethal platinum-DNA adducts

* enable and enhance DNA repair (NER)

* enable cells to tolerate platinum-DNA damage once it occurs

* enhance intracellular detoxification such as the glutathione

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

WHY STEROID ???

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

1. Diversity of possible structureO

HOHO

H

HO

COOHH

OH

OH

estronecholesterol

cholic acid

OH

O

testosterone

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

2. Afinity to cell receptors

androgen receptor

estrogen receptor

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

Reagents as possible ligands

1) Diamine complexes

H2N NH2 N NH2N NH2

HN

COOH

N

COOH

NH2

2) Dicarboxylic complexes

COOHHOOC

R

R = H, NH2, Br

3) Other reagentsOH

OH

S COOH

NH2

OH

OHN

OO

Pt

O PPh3

PPh3

OH

HO

N

PtN

O

O

O

O

O

O

H2NPt

NH3

NH3

Cl

Cl

OH OH

NH6

PtN

ClCl

OH

HO

HO

N

S

S

Pt Cl

Cl

HO

NH2

H2N

PtCl

Cl

Published complexes

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

Synthesis of steroidal amino acid complexes

(i) Boc-L-Met, DCC/benzene; (ii) TFA/CH2Cl2; (iii) K2[PtCl4]/DMF, H2O;

(iv) (Boc)2-L-His, DCC/benzene.

Kvasnica M., Budesinsky M., Swaczynova J., Pouzar V., Kohout L.: Bioorg. Med. Chem. 16 (2008) 3704–3713

HO O

O

NHBoc

O

O

NH2S SO

O

SPt

NH2

Cl Cl

HO O

O

N NHBocBocN

O

O

N NH2HN

O

O

N NH2HN

PtCl Cl

a b c

d e f

(i) (ii) (iii)

(iv) (ii) (iii)

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

RO

H

RO

H

RO

O

O

RO

O

H

H

RO

O OR

O RO

OR

1, R = H1a, R = (Boc)-L-Met1b, R = L-Met1c, R = L-Met-PtCl21d, R = (Boc)2-L-His1e, R = L-His1f, R = L-His-PtCl2




5, R = H5a, R = (Boc)-L-Met5b, R = L-Met5c, R = L-Met-PtCl2




HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

TCS50 (μmol/L) values for complexes 1c-7c and 1f-4f obtained from the Calcein AM assays with the tested cancer and normal cell lines; means ± SD obtained from three independent experiments performed in triplicate.

cell lines (TCS50, μmol/L)

Compound CEM MCF 7 RPMI 8226 A 549 BJ

1c >50 >50 >50 >50 >50

1f >50 >50 >50 >50 >50

2c >50 >50 >50 >50 >50

2f 49.5±0.4 >50 >50 >50 >50

3c 15.5±4.2 44.6±2.6 10.8±2.0 >50 >50

3f 41.8±3.3 >50 46.2±0.5 >50 >50

4c 8.1±3.1 >50 22.2±4.1 43.1±5.6 >50

4f 13.5±1.8 >50 27.4±5.1 >50 >50

5c 14.5±1.2 >50 13.3±2.8 >50 >50

6c 10.1±0.5 >50 46.4±2.9 >50 >50

7c 45.1 >50 47.1 >50 >50

cisplatin 1.6±0.5 9.2±1.4 2.4±0.6 42.4±0.4 5.1±0.2

CEM – T-lymphoblastic leukaemia A549 – human lung adenocarcinoma

MCF7 – human breast adenocarcinoma BJ – human fibroblast

RPMI 8226 – human myeloma


HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

Synthesis of steroidal diamino complexes

(i) Br(CH2)xBr, NaOH/THF, H2O; (ii) 2-(aminomethyl) or 2-(aminoethyl)pyridine/EtOH;

(iii) K2[PtCl4]/DMF, H2O.

HO

(i)

O

O

O

Br x O

O

NH

x

Ny

(ii)

x = 2, 3, 4, 5 x = 2, 3, 4, 5

y = 1, 2

O

O

HN x

(iii)

x = 2, 3, 4, 5

y = 1, 2

N Pt

y

Cl

Cl

HO

(i)

OH

O

OH

Br x O

OH

NH

x

Ny

(ii)

x = 2, 3, 4, 5 x = 2, 3, 4, 5

y = 1, 2

O

OH

HN x

(iii)

x = 2, 3, 4, 5

y = 1, 2

N Pt

y

Cl

Cl

BJ CEM U2OS MCF7(ER+) MCF7 AL(ER+)BT474 (ER+) BT549(ER-) MDA-MB(ER-)MK 81 6.6±0.3 2.4±0.1 9.3±1.0 2.3

MK 97 7.5±0.4 3.0±0.1 13.8±1.1 13.2±3.8 24.7±0.8 12.0±1.7 7.5±1.0 45.1±6.2MK 107 6.6±0.2 2.7±0.4 7.3±0.0 9.7±1.4 15.6±6.2 8.0±0.1 7.4±1.0 22.7±4.4MK 116 9.6±3.9 2.2±0.1 7.5±0.9 9.7±2.0 11.4±4.2 10.1±2.8 6.7±0.8 13.9±1.6MK 85 6.8±0.6 2.5±0.2 7.9±0.8 2.3

MK 98 2.4±0.3 1.7±0.6 5.2±0.4 2.9±0.6 6.0±1.1 3.5±0.1 2.6±0.3 4.0±1.0MK 108 3.1±0.6 1.9±0.3 4.9±0.7 4.2±1.0 6.3±0.9 4.3±0.7 3.9±0.4 5.1±0.5MK 117 2.5±0.1 1.2±0.1 3.0±0.3 3.4±0.6 5.0±1.0 3.1±0.0 2.5±0.3 3.4±1.0MK 89 7.5±0.5 4.3±1.1 23.5±4.9 4.4

MK 101 7.0±3.5 4.9±1.6 8.1±1.6 10.5±2.9 20.9±6.4 14.6±7.1 8.6±1.9 40.1±1.5MK 112 7.6±2.9 3.4±1.5 8.4±0.6 14.8±1.2 30.4±3.5 13.3±6.5 8.5±2.2 31.7±10.8MK 121 9.5±2.9 2.7±0.7 14.7±0.0 10.5±1.5 24.5±15.4 10.0±7.3 7.5±0.6 42.8±0.5MK 90 7.4±0.7 2.2±0 7.1±0.1 3.4

MK 104 3.2±0.2 1.8±0.4 6.3±0.5 4.1±0.9 6.9±0.6 5.0±1.1 3.4±0.1 6.9±0.6MK 113 4.2±1.8 2.0±0.1 7.5±0.6 5.6±1.8 7.2±0.6 3.7±1.1 2.7±0.1 10.5±3.7MK 122 3.0±0.6 1.9±0.2 4.3±1.3 5.0±0.5 5.1±1.3 4.5±1.6 3.3±0.1 13.4±0.6

ESTRADIOL >50 25.2±6.4 32.4±1.6 >50 >50 45.7±3.7 48.4±2.3 21.2±0.3ESTRON >50 35.7±9.7 >50 >50 >50 >50 >50 41.9±4.6

estradiols with 6-membered ring

estrones with 5-membered ring

estrones with 6-membered ring

estradiols with 5-membered ring

BJ – human fibroblast

CEM – T-lymphoblastic leukaemia

U2OS – human osteosarcoma

MCF7, BT474, BT549, MDA-MB - human breast adenocarcinoma

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

Synthesis of steroidal dicarboxylato complexes I

COOHR3

R2R1

H

(i)

R3

R2R1

H

(ii)

CONHR3

R2R1

H

(iii)

COOEt

COOEt

O

O

NO O

CONHR3

R2R1

H OPt

OO

O

NH3

NH3

(i) TSTU, TEA/DMF; (ii) diethyl aminomalonate/DMF;(iii) a) Ba(OH)2/EtOH,H2O

b)[(NH3)2Pt(H2O)2]SO4/EtOH, H2O

R1, R2, R3 = OH – cholic; R1, R2 = OH, R3 = H – chenodeoxycholic; R1, R3 = OH, R2 = H – deoxycholic; R1 = OH, R2, R3 = H – lithocholic; R1, R2, R3 = H – cholanic

1) K2[PtCl4] + 4 KI → K2[PtI4] + 4 KCl

2) K2[PtI4] + 2 NH3 → (NH3)2PtI2 + 2 KI

3) (NH3)2PtI2 + Ag2SO4 → [(NH3)2Pt(H2O)2]SO4 + AgI

4) [(NH3)2Pt(H2O)2]SO4 + BaX → (NH3)2PtX + BaSO4

Synthesis of platinum reagents for dicarboxylato complexes

Synthesis of cisplatin

Source: wikipedia

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

Synthesis of steroidal dicarboxylato complexes II

1) Knoevenagel condensation

O

H

CH2(CO2Me)2, TiCl4

pyridine, THF

H

MeO2C

CO2Me

75 %

H

MeO2C

CO2Me

10 %

HH

H

O

H

H

10 %H

H

CH2(CO2Me)2, TiCl4

pyridine, THF

25 %MeOMeO

O CO2MeMeO2C

2) Substitution of tosylates

TosO

H

NaCH(CO2Me)2,

toluene

H

MeO2C

CO2Me

86 %

HH

NaCH(CO2Me)2

xylene

MeO

OTos

no reaction

O

HO

NH2

Pt

H2N

HO

Cl

Cl

HO

NH2

Pt

H2N

Cl

Cl

Platinum in steroidal skeleton

HO

COOHH

OH

OH

O

O

H

HO

H

OH

O

O

HO

Thank you for your attention.

steroidal platinum complexes

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