HO COOH H 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
Jan 12, 2016
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
2, R = H2a, R = (Boc)-L-Met2b, R = L-Met2c, R = L-Met-PtCl22d, R = (Boc)2-L-His2e, R = L-His2f, R = L-His-PtCl2
3, R = H3a, R = (Boc)-L-Met3b, R = L-Met3c, R = L-Met-PtCl23d, R = (Boc)2-L-His3e, R = L-His3f, R = L-His-PtCl2
4, R = H4a, R = (Boc)-L-Met4b, R = L-Met4c, R = L-Met-PtCl24d, R = (Boc)2-L-His4e, R = L-His4f, R = L-His-PtCl2
5, R = H5a, R = (Boc)-L-Met5b, R = L-Met5c, R = L-Met-PtCl2
6, R = H6a, R = (Boc)-L-Met6b, R = L-Met6c, R = L-Met-PtCl2
7, R = H7a, R = (Boc)-L-Met7b, R = L-Met7c, R = L-Met-PtCl2
Kvasnica M., Budesinsky M., Swaczynova J., Pouzar V., Kohout L.: Bioorg. Med. Chem. 16 (2008) 3704–3713
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
Kvasnica M., Budesinsky M., Swaczynova J., Pouzar V., Kohout L.: Bioorg. Med. Chem. 16 (2008) 3704–3713
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.