Intermezzo: metals in life

MST - ANO 1
 Metals in Life

• Bioinorganic chemistry
• Role of essential elements (including uptake from food)
• Role of toxic elements
• Metals as medicine

MST - ANO 2
 Bioinorganic Chemistry of Alcohol

• Intake of EtOH: “Happy”

Zn in Alcohol dehydrogenase

• Formation of Acetaldehyde: “Hangover”

Mo in Aldehyde Oxidase

• Formation of Acetic Acid:

The Recovery Stage

disulfiram; antabuse

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 DAH of trace elements

dagelijks aanbevolen hoeveelheid

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 Metal supplements

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 Li2CO3 manic
depression
111In-dtpafor
brain imaging
67Ga-citrate for
153Sm for pain diagnosis
relieve in bone
cancer Ag salts for
skin protection
Gd-bopta for
MRI imaging
99mTc-cardiolite

for hart scan

Bi-citrate for
Auranofin for
ulcer healing
arthritis
BaSO4 as a
Carboplatin and
contrast agent
cisplatin for
for X-ray scan
anticancer
MST - ANO 6
 Organic vs Inorganic Drugs

• Easy one-electron reductions possible with many metal ions

• Easy changes in structure due to metal-ligand dissociation
reactions:
– use as pro-drugs;
– kinetic differences are metal dependent
• Metal ions as such are non-degradable and have to be excreted.

MST - ANO 7
 Statistical Information

• Each year in NL: 180,000 deaths, of which 25% due to cancer

(mostly elderly people)
• Half of the new patients (60,000/y) will undergo chemotherapy
• 25% of these is treated with cisplatin (mostly injections)
• Cisplatin may cure up to 70%; some tumors above 90%

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 Cis-platin chemotherapy

Cl NH3
Pt
Cl NH3

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 Clinical inconveniences

• Severe side effects:

–nephrotoxicity, vomiting, deafness
–Appearance of cisplatin-resistant tumor cells
–Several tumor lines are less (not) sensitive

Binding takes places on DNA, and that

leads to cell killing.
QUESTION: How does it reach the DNA,
and where does it go afterwards??

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 Questions from chemistry

• Why cisPt active, transPt not?

• Can we make more active Pt complexes
• Can we understand the side effects?
• What happens with cisPt in blood
• Can it be orally administered as pills?
• How does cisPt enter the tumor cells?
• Why are tumor cells affected and normal cells not?

MST - ANO 11
 Transport of Pt in the body

Oral Drugs
Injection Pt 1997

Pt enters all cells

Some Pt expelled
Transport

diuretic Apoptosis CELL NUCLEUS

DNA binding
Kidney (toxicity) LIVER HMG binding

Excretion: 50% < 48hrs; rest < 2 months

MST - ANO HMG = High-mobility group protein 12

 Various ligands on Pt

In blood [Cl–] = 100 mM; cis-[PtCl2(NH3)2]

Neutral complex can enter the cell

In cell [Cl–] = 4 mM; hydrolysis: cis-[Pt(H2O)2(NH3)2]2+

Charged complex can enter the nucleus

Is attracted to DNA

Complication: Pt also binds to S (proteins)

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 Cisplatin Research

Major problems:
Side effects and Resistance Development

EVALUATION
TESTS

Mechanistic Research NEW DRUGS

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 Four newer Pt compounds

O
OAc
NH3 Cl
O O Pt
O Pt NH3 C6H11NH2 Cl
OAc
NH3
CBDCA JM-216(oral)

O O
Cl
NH2
Pt Cl Pt N
NH2 O O NH3
Me
Oxaliplatin AMD-473
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 Double helix in DNA

Binding sites for metals:

• O atoms (red)
hard metals
• N atoms (blue)
soft metals

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 NH2
DNA
N C
1
N O
HO 5'
O O
H 3' 2'
H H3 C
H H NH
O H
1
T
N O
O P O-
O 5'
O O
G
Possible binding sites H 3' 2'
H
N
H H 7 1NH
O H
3
O P O- N N NH2
O 5'
O NH2
H 3' 2' H
d(CpTpGpA) N
H H 7 1N
O H
3
O P O- N N H
O 5'
O A
H 3' 2'H
H H
MST - ANO OH H 17
 Nucleic acid bases and base pairs

N7 not involved in H-bonding  is available for Pt binding;

(N3: sterically protected)

7 7

3 3

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 Hypothesis mechanism cisPt

• Cisplatin binds to DNA at a specific position: N7-positions of two

neighboring guanine bases
• The resulting change in the DNA is rather small (kink); replication of
platinated DNA in cells is not possible
• This change is not recognized in (some) tumor cells
• In other (healthy, resistant) cells the damage is recognized and
removed

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 Most frequent Pt-DNA adducts

H3N NH3 H3N NH3

Pt Pt
G G A G
C C T C
1,2-d(GG) intrastrand 1,2-d(AG) intrastrand
(60-65 %) (20-25 %)

H3N NH3 H3N

Pt NH3
Pt
G X G G C
C X C C G
1,3-d(GXG) intrastrand 1,2-d(GG) interstrand
(~2 %) (1.5 %)
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 Most frequent binding on DNA

H3N Cl G G-N7
H3N
Pt + Pt
H3N
H3N G-N7
Cl
G
Cisplatin

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 1,2 (GG) intrastrand structure

7 NMR structures
2 X-ray structures (high res)
1 X-ray complexed with HMG

•Pt in major groove

•Duplex is bent toward major groove
•Duplex is unwound
•minor groove opens up

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 DNA + HMG protein

High-mobility group protein

Structure of HMG protein

at
1,2 intrastrand GG adduct Pt

possibly blocking repair enzymes

MST - ANO S.J. Lippard, Nature, 1999 23

 Ru anti-tumor complexes

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 Why are metallodrugs toxic ?
Sylvestre Bonnet, MCBIM

[Cl-] 100 mM

[Cl-] 5 mM

DNA
M
M Cl M Cl
proteins
M Cl M OH2
M OH2 M
Spontaneous

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 Controlling Ru aquation

37 °C
2+ 2+
+ L
water
in the dark

% Ru-OH2 formation

Time /
4h 3 days 3 weeks
L
Cl- 90 90 90

N-acetylmethionine 0 0 0

biotin 0 3 30

MST - ANO Bonnet et al, Chem. Eur. J. 2011 p9924 26

 Controlling Ru aquation

visible light
2+ 2+
+ L
water
r.t.

Ru-OH2 L = biotin
Ru-S

λ = 450 nm

MST - ANO Bonnet et al, Chem. Eur. J. 2011 p9924 27

 Aim of the research

[Cl-] 100 mM

[Cl-] 5 mM

Ru

Ru S Ru S

Ru S Ru OH2
Ru

Light
irradiation

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 Photo-Activated Chemo-Therapy

Ru-S
Ru-S Ru-OH2

• new Ru compounds for photochemotherapy

• low toxicity in the dark
• red light activation

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 Synthesized complexes

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 [Ru(tpy)(qipy)(D-biotin)]Cl2

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 Concluding Remarks

• Metal compounds can be very important in Medicine. Cisplatin

and derivatives are great drugs, but still poorly understood
from a pharmacokinetics point of view; stay in nucleus may be
relatively short;
• Specific binding at G-N7 sites may be the explanation why S
binding is at best temporarily;
• New photoactivatable compounds might give better selectivity
and less side-effects

MST - ANO 32